Antitumoral analogs of et-743

ABSTRACT

Antitumour compounds have the five membered fused ring ecteinascidin structure of the formula (XIV). The present compounds lack a 1,4-bridging group as found in the ecteinascidins. They have at the C-1 position a substituent selected from an optionally protected or derivatised aminomethylene group or an optionally protected or derivatised hydroxymethylene group.

The present invention relates to antitumoral compounds, and inparticular to antitumoral analogs of ecteinascidin 743, ET-743.

BACKGROUND OF THE INVENTION

European Patent 309,477 relates to ecteinascidins 729, 743, 745, 759A.759B and 770. The ecteinascidin compounds are disclosed to haveantibacterial and other useful properties. Ecteinascidin 743 is nowundergoing clinical trials as an antitumour agent.

Ecteinascidin 743 has a complex tris(tetrahydroisoquinolinephenol)structure of the following formula (I):

In ecteinascidin 743, the 1,4 bridge has the structure of formula (IV):

Other known ecteinascidins include compounds with a different bridgedcyclic ring system such as occurs in ecteinascidin 722 and 736, wherethe bridge has the structure of formula (V):

ecteinascidins 583 and 597, where the bridge has the structure offormula (VI):

and ecteinascidin 594 and 596, where the bridge has the structure offormula (VII):

The complete structure for these and related compounds is given in J.Am. Chem. Soc. (1996) 118, 9017–9023. This article is incorporated byreference.

The ecteinascidins are currently prepared by isolation from extracts ofthe marine tunicate Ecteinascidiii turbinata. The yield is low, andalternative preparative processes have been sought.

A synthetic process for producing ecteinascidin compounds is describedin U.S. Pat. No. 5,721,362, see also WO 9812198. The claimed method islong and complicated. By way of illustration, there are 38 Examples eachdescribing one or more steps in the synthetic sequence to arrive atecteinascidin 743.

Claim 25 of U.S. Pat. No. 5,721,362 is directed at an intermediatephenol compound of a given formula (11), which we refer to also asIntermediate 11 or Int-11. It has the followingbis(tetrahydroisoquinolinephenol) structure (II):

where MOM is a methoxymethyl substituent and TBDPS is atert-butyldiphenylsilyl substituent.

From Intermediate 11 it is possible to synthesise another interestingantitumour agent, phthalascidin, see Proc. Natl. Acad. Sci. USA, 96,3496–3501, 1999. Phthalascidin is a bis(tetrahydroisoquinolinephenol)derivative of formula (III):

More generally, phthalascidin and related compounds are described in WO0018233. Claim 1 is directed at compounds of formula:

-   wherein the substituent groups defined by R₁, R₂, R₃, R₅, R₆, R₇, R₈    and R₉ are each independently selected from the group consisting of    H, OH, OR′, SH, SR′, SOR′, SO₂R′, NO₂, NH₂, NHR′, N(R′)₂, NHC(O)R′,    CN, halogen, ═O, C(═O)H, C(═O)R′, CO₂H, CO₂R′, C₁–C₁₂ alkyl, C₂–C₁₂    alkenyl, C₂–C₁₂ alkynyl, substituted or unsubstituted aryl,    substituted or unsubstituted aralkyl, and substituted or    unsubstituted heteroaromatic;-   wherein each of the R′ groups is independently selected from the    group consisting of H, OH, NO₂, NH₂, SH, CN, halogen, ═O, C(═O)H,    C(═O)CH₃, CO₂H, CO₂CH₃, C₁–C₁₂ alkyl, C₂–C₁₂ alkenyl, C₂–C₁₂    alkynyl, aryl, aralkyl, and heteroaromatic;-   wherein each dotted circle represents one, two or three optional    double bonds;-   wherein R₇ and R₈ may be joined into a carbocyclic or heterocyclic    ring system;-   and wherein X₁ and X₂ are each independently defined as above for    R₁–R₈ and further include various permitted definitions.

Further naturally occuring compounds are known which lack a bridgedcyclic ring system. They include the bis(tetrahydroisoquinolinequinone)antitumor-antimicrobial antibiotics safracins and saframycins and themarine natural products renieramicins and xestomycin isolated fromcultured microbes or sponges. They all have a common dimerictetrahydroisoquinoline carbon framework. These compounds can beclassified into four types, types I to IV, with respect to the oxidationpattern of the aromatic rings.

Type I, dimeric isoquinolinequinones, is a system of formula (VIII) mostcommonly occurring in this class of compounds, see the following tableI.

TABLE I Structure of Type I Saframycin Antibiotics.

Substituents Compound R^(14a) R^(14b) R²¹ R^(25a) R^(25b) R^(25c)saframycin A H H CN O O CH₃ saframycin B H H H O O CH₃ saframycin C HOCH₃ H O O CH₃ saframycin G H OH CN O O CH₃ saframycin H H H CN OHCH₂COCH₃ CH₃ saframycin S H H OH O O CH₃ saframycin Y₃ H H CN NH₂ H CH₃saframycin Yd₁ H H CN NH₂ H C₂H₅ saframycin Ad₁ H H CN O O C₂H₅saframycin Yd₂ H H CN NH₂ H H saframycin Y_(2b) H Q^(b) CN NH₂ H CH₃saframycin Y_(2b−d) H Q^(b) CN NH₂ H C₂H₅ saframycin AH₂ H H CN H^(a)OH^(a) CH₃ saframycin AH₂Ac H H CN H OAc CH₃ saframycin AH₁ H H CNOH^(a) H^(a) CH₃ saframycin AH₁Ac H H CN OAc H CH₃ saframycin AR₃ H H HH OH CH₃ ^(a)assignments are interchangeable. ^(b)where the group Q isof formula (IX):

Type I aromatic rings are seen in saframycins A, B and C; G and H; and Sisolated from Streptomyces lavendulae as minor components. A cyanoderivative of saframycin A, called cyanoquinonamine, is known fromJapanese Kokai JP-A2 59/225189 and 60/084,288. Saframycins Y₃, Yd₁, Ad₁,and Yd₂ were produced by S. lavenidulae by directed biosynthesis, withappropriate supplementation of the culture medium. Saframycins Y_(2b)and Y_(2b-d) dimers formed by linking the nitrogen on the C-25 of oneunit to the C-14 of the other, have also been produced in supplementedculture media of S. lavendulae. Saframycins AR, (=AH₂,), a microbialreduction product of saframycin A at C-25 produced by Rhodococcusamidophilus, is also prepared by nonstereoselective chemical reductionof saframycin A by sodium borohydride as a 1:1 mixture of epimersfollowed by chromatographic separation [the other isomer AH₁ is lesspolar]. The further reduction product saframycin AR₃,21-decyano-25-dihydro-saframycin A. (=25-dihydrosaframycin B) wasproduced by the same microbial conversion. Another type of microbialconversion of saframycin A using a Nocardia species produced saframycinB and further reduction by a Mycobacterium species produced saframycinAH¹Ac. The 25-O-acetates of saframycin AH₂ and AH₁ have also beenprepared chemically for biological studies.

Type I compounds of formula (X) have also been isolated from marinessponges, see Table II.

TABLE II Structures of Type I Compounds from Marine Sponges.

Substituents R^(14a) R^(14b) R²¹ R renieramycin A OH H H —C(CH₃)═CH—CH₃renieramycin B OC₂H₅ H H —C(CH₃)═CH—CH₃ renieramycin C OH O O—C(CH₃)═CH—CH₃ renieramycin D OC₂H₅ O O —C(CH₃)═CH—CH₃ renieramycin E HH OH —C(CH₃)═CH—CH₃ renieramycin F OCH₃ H OH —C(CH₃)═CH—CH₃ xestomycinOCH₃ H H —CH₃

Renieramycins A–D were isolated from the antimicrobial extract of asponge, a Reniera species collected in Mexico, along with thebiogenetically related monomeric isoquinolines renierone and relatedcompounds. The structure of renieramycin A was initially assigned withinverted stereochemistry at C-3, C-11, and C-13. However, carefulexamination of the ¹H NMR data for new, related compounds renieramycinsE and F, isolated from the same sponge collected in Palau, revealed thatthe ring junction of renieramycins was identical to that of saframycins.This result led to the conclusion that the formerly assignedstereochemistry of renieramycins A to D must be the same as that ofsaframycins.

Xestomycin was found in a sponge, a Xestosponigia species collected fromSri Lancan waters.

Type II compounds of formula (XI) with a reduced hydroquinone ringinclude saframycins D and F, isolated from S. lavendulae, andsaframycins Mx-1 and Mx-2, isolated from Myxococcus xanthlus. See tableIII.

TABLE III Type II Compounds

Substituents Compound R^(14a) R^(14b) R²¹ R^(25a) R^(25a) R^(25c)saframycin D O O H O O CH₃ saframycin F O O CN O O CH₃ saframycin Mx-1 HOCH₃ OH H CH₃ NH₂ saframycin Mx-2 H OCH₃ H H CH₃ NH₂

The type III skeleton is found in the antibiotics safracins A and B,isolated from cultured Pseudomonas fluorescens. These antibiotics offormula (XII) consist of a tetrahydroisoquinoline-quinone subunit and atetrahydroisoquninolinephenol subunit.

where R²¹ is —H in safracin A and is —OH in safracin B.

Saframycin R, the only compound classified as the Type IV skeleton, wasalso isolated from S. lavendulae. This compound of formula (XIII),consisting of a hydroquinone ring with a glycolic ester sidechain on oneof the phenolic oxygens, is conceivably a pro-drug of saframycin Abecause of its moderate toxicity.

All these known compounds have a fused system of five rings (A) to (E)as shown in the following structure of formula (XIV):

The rings A and E are phenolic in the ecteinascidins and some othercompounds, while in other compounds, notably the saframycins, the ringsA and E are quinolic. In the known compounds, the rings B and D aretetrahydro, while ring C is perhydro.

SUMMARY OF THE INVENTION

The present invention provides new compounds with the fused system offive rings (A) to (E). In particular, it provides new compounds whichcan be made from intermediates described in WO 9812198 or by a newprocess which is part of this invention. In this latter respect, werefer to our WO 0069862 published 23 Nov. 2000, and which relates tohemisynthetic methods and new compounds. The present application claimspriority from that PCT filing, and we incorporate that text by referenceto the extent that there is disclosure therein which is not in thepresent specification.

In WO 0069862, various routes are described for the preparation ofecteinascidin compounds, including ecteinascidin 743, as well asecteinascidin analogs including phthaliscidin. The present invention isfounded partly on the use of intermediates of WO 0069862 to preparefurther analogs of the ecteinasacidins.

Preferred Embodiments

We have found that compounds of the invention have exceptional activityin the treatment of cancers, such as leukaemias, lung cancer, coloncancer, kidney cancer and melanoma.

Thus, the present invention provides a method of treating any mammal,notably a human, affected by cancer which comprises administering to theaffected individual a therapeutically effective amount of a compound ofthe invention, or a pharmaceutical composition thereof.

The present invention also relates to pharmaceutical preparations, whichcontain as active ingredient a compound or compounds of the invention,as well as the processes for their preparation.

Examples of pharmaceutical compositions include any solid (tablets,pills, capsules, granules, etc.) or liquid (solutions, suspensions oremulsions) with suitable composition or oral, topical or parenteraladministration, and they may contain the pure compound or in combinationwith any carrier or other pharmacologically active compounds. Thesecompositions may need to be sterile when administered parenterally.

Administration of the compounds or compositions of the present inventionmay be by any suitable method, such as intravenous infusion, oralpreparations, intraperitoneal and intravenous administration. We preferthat infusion times of up to 24 hours are used, more preferably 2–12hours, with 2–6 hours most preferred. Short infusion times which allowtreatment to be carried out without an overnight stay in hospital areespecially desirable. However, infusion may be 12 to 24 hours or evenlonger if required. Infusion may be carried out at suitable intervals ofsay 2 to 4 weeks. Pharmaceutical compositions containing compounds ofthe invention may be delivered by liposome or nanosphere encapsulation,in sustained release formulations or by other standard delivery means.

The correct dosage of the compounds will vary according to theparticular formulation, the mode of application, and the particularsitus, host and tumour being treated. Other factors like age, bodyweight, sex, diet, time of administration, rate of excretion, conditionof the host, drug combinations, reaction sensitivities and severity ofthe disease shall be taken into account. Administration can be carriedout continuously or periodically within the maximum tolerated dose.

The compounds and compositions of this invention may be used with otherdrugs to provide a combination therapy. The other drugs may form part ofthe same composition, or be provided as a separate composition foradministration at the same time or a different time. The identity of theother drug is not particularly limited, and suitable candidates include:

-   a) drugs with antimitotic effects, especially those which target    cytoskeletal elements, including microtubule modulators such as    taxane drugs (such as taxol, paclitaxel, taxotere, docetaxel),    podophylotoxins or vinca alkaloids (vincristine, vinblastine):-   b) antimetabolite drugs such as 5-fluorouracil, cytarabine,    gemcitabine, purine analogues such as pentostatin, methotrexate);-   c) alkylating agents such as nitrogen mustards (such as    cyclophosphamide or ifosphamide);-   d) drugs which target DNA such as the antracycline drugs adriamycin,    doxorubicin, pharmorubicin or epirubicin;-   e) drugs which target topoisomerases such as etoposide;-   f) hormones and hormone agonists or antagonists such as estrogens,    antiestrogens (tamoxifen and related compounds) and androgens,    flutamide, leuprorelin, goserelin, cyprotrone or octreotide;-   g) drugs which target signal transduction in tumour cells including    antibody derivatives such as herceptin;-   h) alkylating drugs such as platinum drugs (cis-platin,    carbonplatin, oxaliplatin, paraplatin) or nitrosoureas;-   i) drugs potentially affecting metastasis of tumours such as matrix    metalloproteinase inhibitors;-   j) gene therapy and antisense agents;-   k) antibody therapeutics;-   l) other bioactive compounds of marine origin, notably the didemnins    such as aplidine;-   m) steroid analogues, in particular dexamethasone;-   n) anti-inflammatory drugs, in particular dexamethasone;-   o) anti-emetic drugs, in particular dexamethasone;-   p) skeletal muscle protectors, such as L-carnitine or precursor    amino acids.

The present invention also extends to the compounds of the invention foruse in a method of treatment, and to the use of the compounds in thepreparation of a composition for treatment of cancer.

In one aspect of the invention, we make no claim to the compounds 2.3.5. 8-OH-2. and 14 to 21 described in one or more of the GB prioritypatent applications for our PCT application published as 0069862. In arelated aspect, the present invention extends to compounds which differin respect of one or more of the substituents present at C-1, C-5, C-7,C-8, or C-18 in the compounds of these GB priority patent applications.

The compounds of this invention include compounds which do not have ahydroxy group at the C-18 position. Furthermore, the compounds of thisinvention include compounds which do not have a dicarboximidomethylsubstituent, such as phthalimidomethyl, at the C-1 position. Inparticular, we provide active compounds where the subsituent X₁ is notas shown in the penultimate line at page 19 of WO0018233.

In one aspect, the analogs of this invention are typically of theformula (XVIIa):

where

-   R¹ is an optionally protected or derivatised aminomethylene group,    an optionally protected or derivatised hydroxymethylene group;-   R⁴ is —H;-   R⁵ is —H or —OH;-   R⁷ is —OCH₃ and R⁸ is —OH or R⁷ and R⁸ together form a group —O—CH,    —O—,-   R^(14a) and R^(14b) are both —H or one is —H and the other is —OH,    —OCH₃ or —OCH₂CH₃, or R^(14a) and R^(14b) together form a keto    group; and-   R¹⁵ is —H or —OH;-   R²¹ is —H, —OH or —CN;    and derivatives including acyl derivatives thereof especially where    R⁵ is acetyloxy or other acyloxy group of up to 4 carbon atoms.

In the present invention, a key class of products includes phthalascidinand has the general formula (XX):

where R¹ is an amidomethylene group; R⁵ is a small oxy-sidechain; andR²¹ is a cyano group or a hydroxy group. For phthalascidin, R¹ is aphthalimidomethylene group; R⁵ an acetoxy group; and R²¹ is a cyanogroup. Other groups for R¹ include mono- and di-N-substitutedamidomethylenes as well as other cyclic amidomethylenes, and othergroups for R⁵ include further C₁–C₄ acyl groups, as well as C₁–C₄ alkylgroups.

In the present invention, a key class of intermediates and analogsincludes Intermediate 11 and has the general formula (XXI):

where Prot¹ and Prot² are hydroxy protecting groups, preferablydifferent. For Intermediate 11 itself, the group Prot¹ is amethoxymethyl group, and Prot² is a t-butyldiphenylsilyl group.

In the light of the preceding explanations, it can be seen that thepresent invention provides novel analogs and novel intermediatecompounds. Depending on ring A, the compounds include those of formula(XXIIa):

where:

-   R¹ is —CH₂NH₂ or —CH₂OH, or a protected or derivatised version of    such a group and R⁴ is —H;-   R⁵ is —OH or a protected or derivatised version of such a group;-   R^(14a) and R^(14b) are both —H or one is —H and the other is —OH or    a protected or derivatised version of such a group, —OCH₃ or    —OCH₂CH₃, or R^(14a) and R^(14b) together form a keto group;-   R¹² is —H—, —CH₃— or —CH₂CH₃—;-   R¹⁵ is —H, —OH or a protected or derivatised version of such a    group; and-   R¹⁸ is —OH or a protected or derivatised version of such a group.

In one embodiment, preferably at least of R¹, R⁵, R^(14a), R^(14b), R¹⁵or R¹⁸ is a protected or derivatised group.

In one variation of this invention, the group R¹ is not atert-butyldiphenylsilyl substituent and/or the group R¹⁸ is not amethoxymethyloxy group.

Preferably R¹ is —CH₂NH₂ or —CH₂OH, or a protected or derivatisedversion of such a group and R⁴ is —H.

Preferably R^(14a) and R^(14b) are both —H.

Preferably R¹² is —CH₃.

One preferred class of intermediates includes the compound which weidentify as compound 25, of formula:

The preferred class is thus of the general formula where the group MOMis replaced by any other protecting group, and/or the allyl is replacedby any other protecting group.

Other preferred intermediates includes the compounds which we identifyas compounds 17, 43 and 45.

Other N-acyl derivatives may readily be made from compound 45 and are animportant part of this invention. Suitable acyl groups include thosepreviously mentioned. The corresponding 21-hydroxy compounds are alsouseful and are among the active compounds which we have found.

From the activity data and other considerations, it can be seen that theactive compounds of this invention include a preferred class ofcompounds of the general formula (XXIII):

where R¹ is as previously defined for formula (XVIIb) and is preferablya derivatised aminomethylene group of moderate bulk;

R⁵ is as previously defined for formula (XVIIb) and is preferably aderivatised hydroxy group of low bulk;

R¹² is as previously defined and is preferably —CH₃—; and

R²¹ is a hydroxy or cyano group.

R¹ is suitably a hydrophobic group and which thus lacks free amino,hydroxy or other hydrophilic function. Typically R¹ is a group—CH₂—NH₂—CO—R^(a), where R^(a) is as defined but preferably has a linearchain length of less than 20 atoms, more preferably less than 15 or 10atoms, where a 1,4-phenyl is counted as a chain length of four atoms andsimilar considerations apply to other cyclic groups (for example,1,2-cyclohexyl is chain length of two), and the linear chain of lessthan 10, 15 or 20 atoms can itself be substituted. In particular, thedata suggests there is a balance to be achieved between having no suchgroup R^(a)—CO— and having a large, bulky group.

In one variation, we prefer that R¹ is free from cyclic groups,especially aromatic groups. In a related variation, the presentinvention does not prepare the compounds which are described in thearticle Proc. Natl. Acad. Sci. USA, 96, 3496–3501, 1999, incorporated byreference. Our preferred groups for R¹ exclude the correspondingsubstituents CH₂R₂ shown in Table 1 of that article, specifically thegroups A, B, C and D for R₂.

R⁵ is preferably an acetyl group.

In particularly preferred compounds, the group R¹ is acylated on an —NH₂group, and for example N-acyl derivatives can be formed from groups—CH₂NH, and —CH₂—NH-aa. The acyl derivatives can be N-acyl or N-thioacylderivatives thereof. The acyl groups can be of formula —CO—R^(a), whereR^(a) is as defined and is chosen to meet the indicated criteria.Suitable acyl groups include alanyl, arginyl, aspartyl, asparagyl,cystyl, glutamyl, glutaminyl, glycyl, histidyl, hydroxyprolyl,isoleucyl, leucyl, lysyl, methionyl, phenylalanyl, prolyl, seryl,threonyl, thyronyl, tryptophyl, tyrosyl, valyl, as well as other aminoacid acyl groups, which may be L- or D-. Such amino acid acyl groups arepreferred derivatised on the amino group to give hydrophobicity.

In a variation, the group R¹ is a derivatised hydroxymethylene group.Similar considerations apply as with the derivatised aminomethylenegroup.

The invention extends to compounds where the various substituents aroundthe ring are as defined in the WO 0018233, which we incorporate byreference. Thus, as appropriate, substituents in the present compoundscan be chosen, among other possibilites from H, OH, OR′, SH, SR′, SOR′,SO₂R′, NO₂, NH₂, NHR′, N(R′)₂, NHC(O)R′, CN, halogen, ═O, C₁–C₆ alkyl,substituted or unsubstituted aryl, substituted or unsubstituted aralkyl,and substituted or unsubstituted heteroaromatic;

wherein each of the R′ groups is independently selected from the groupconsisting of H, OH, NO₂, NH₂, SH, CN, halogen, ═O, C(═O)H, C(═O)CH₃,CO₂H, CO₂CH₃, C₁–C₆ alkyl, phenyl, benzyl and heteroaromatic.

Suitable halogen substituents in the compounds of the present inventioninclude F. Cl, Br and I.

Alkyl groups preferably have from 1 to about 12 carbon atoms, morepreferably 1 to about 8 carbon atoms, still more preferably 1 to about 6carbon atoms, and most prefereably 1, 2, 3 or 4 carbon atoms. Methyl,ethyl and propyl including isopropyl are particularly preferred alkylgroups in the compounds of the present invention. As used herein, theterm alkyl, unless otherwise modified, refers to both cyclic andnoncyclic groups, although cyclic groups will comprise at least threecarbon ring members.

Preferred alkenyl and alkynyl groups in the compounds of the presentinvention have one or more unsaturated linkages and from 2 to about 12carbon atoms, more preferably 2 to about 8 carbon atoms, still moreprefereably 2 to about 6 carbon atoms, even more prefereably 1, 2, 3 or4 carbon atoms. The terms alkenyl and alkynyl as used herein refere toboth cyclic and noncyclic groups, although straight or branchednoncyclic groups are generally more preferred.

Preferred alkoxy groups in the compounds of the present inventioninclude groups having one or more oxygem linkages and from 1 to about 12carbon atoms, more preferably from 1 to about 8 carbon atoms, and stillmore preferably 1 to about 6 carbon atoms, and most preferably 1, 2, 3or 4 carbon atoms.

Preferred alkylthio groups in the compounds of the present inventionhave one or more thioether linkages and from 1 to about 12 carbon atoms,more prefereably from 1 to about 8 carbon atoms, and still morepreferably 1 to about 6 carbon atoms. Alkylthio groups having 1, 2, 3 or4 carbon atoms are particularly preferred.

Preferred alkylsulfinyl groups in the compounds of the present inventioninclude those groups having one or more sulfoxide (SO) groups and from 1to about 12 carbon atoms, more preferably from 1 to about 8 carbonatoms, and still more preferably 1 to about 6 carbon atoms.Alkylsulfinyl groups having 1, 2, 3 or 4 carbon atoms are particularlypreferred.

Preferred alkylsulfonyl groups in the compounds of the present inventioninclude those groups having one or more sulfonyl (SO₂) groups and from 1to about 12 carbon atoms, more preferably from 1 to about 8 carbonatoms, and still more preferably 1 to about 6 carbon atoms.Alkylsulfonyl groups having 1, 2, 3 or 4 carbon atoms are particularlypreferred.

Preferred aminoalkyl groups include those groups having one or moreprimary, secondary and/or tertiary amine groups, and from 1 to about 12carbon atoms, more preferably 1 to about 8 carbon atoms, still morepreferably I to about 6 carbon atoms, even more preferably 1, 2, 3 or 4carbon atoms. Secondary and tertiary amine groups are generally morepreferred than primary amine moieties.

Suitable heteroaromatic groups in the compounds of the present inventioncontain one, two or three heteroatoms selected from N, O or S atoms andinclude, e.g., coumarinyl including 8-coumarinyl, quinolinyl including8-quinolinyl, pyridyl, pyrazinyl, pyrimidyl, furyl, pyrrolyl, thienyl,thiazolyl, oxazolyl, imidazolyl, indolyl, benzofuranyl and benzothiazol.Suitable heteroalicyclic groups in the compounds of the presentinvention contain one, two or three heteroatoms selected from N, O or Satoms and include, e.g., tetrahydrofuranyl, tetrahydropyranyl,piperidinyl, morpholino and pyrrolindinyl groups.

Suitable carbocyclic aryl groups in the compounds of the presentinvention include single and multiple ring compounds, including multiplering compounds that contain separate and/or fused aryl groups. Typicalcarbocyclic aryl groups contain 1 to 3 separate or fused rings and from6 to about 18 carbon ring atoms. Specifically preferred carbocyclicarykl groups include phenyl including substituted phenyl, such as2-substituted phenyl 3-substituted phenyl, 2,3-substituted phenyl,2,5-substituted phenyl, 2,3,5-substituted and 2,4,5-substituted phenyl,including where one or more of the phenyl substituents is anelectron-withdrawing group such as halogen, cyano, nitro, alkanoyl,sulfinyl, sulfonyl and the like; naphthyl including 1-naphthyl and2-naphthyl; biphenyl; phenanthryl; and anthracyl.

Any references herein to substituted groups in the compounds of thepresent invention refer to the specified moiety that may be substitutedat one or more available positions by one or more suitable groups, e.g.,halogen such as fluoro, chloro, bromo and iodide; cyano; hydroxyl;nitro; azido; alkanoyl such as a C1–6 alkanoyl group such as acyl andthe like; carboxamido; alkyl groups including those groups having 1 toabout °2 carbon atoms or from 1 to about 6 carbon atoms and morepreferably 1–3 carbon atoms; alkenyl and alkynyl groups including groupshaving one or more unsaturated linkages and from 2 to about 12 carbon orfrom 2 to about 6 carbon atoms; alkoxy groups having those having one ormore oxygen linkages and from 1 to about 12 carbon atoms or 1 to about 6carbon atoms; aryloxy such as phenoxy; alkylthio groups including thosemoieties having one or more thioether linkages and from 1 to about 12carbon atoms or from 1 to about 6 carbo atoms; alkylsulfinyl groupsincluding those moieties having one or more sulfinyl linkages and from 1to about 12 carbon atoms or from 1 to about 6 carbon atoms;alkylsulfinyl groups including those moieties having one or moresulfonyl linkages and from 1 to about 12 carbon atoms or from 1 to about6 carbon atoms; aminoalkyl groups such as groups having one or more Natoms and from 1 to about 12 carbon atoms or from 1 to about 6 carbonatoms; carbocyclic aryl having 6 or more carbons, particularly phenyl(e.g., R being a substituted or unsubstituted biphenyl moiety); andaralkyl such as benzyl.

Without being exhaustive, in terms of the formula:

preferred compounds of this invention have one or more of the followingdefinitions:

-   R₁ is —OR, where R is H, acyl, especially acetyl, alkyl-CO— (alkyl    being up to about 20 carbon atoms, more preferably from 1 to about    12 carbon atoms, and especially an odd number of carbon atoms such    as 3, 5, 7 and 9), cycloalkyl-alkyl-CO— and especially alkyl    groupings with a terminal cyclohexyl group and up to six additional    carbon atoms in the sidechain, or a protecting group, especially    methoxymethyl, and R₁ is more especially OH.-   R₂ is methoxy.-   R₃ is methyl.-   R₄ is hydrogen.-   R₅ is methyl or hydrogen, especially methyl.-   R₆ is —CN or —OH.-   X₁ is —NHR′, —NH-aa-R′ or —OR′ where aa is an optionally protected    amino acid acyl group, especially alanine, phenylalanine, cysteine,    proline, valine, arginine, tryptophan or other amino acid. Other    possibilities for X₁ include —N(R′)₂, —N(R′)-aa-R′, and —N-(aa-R′)₂.    In the case of any group -aa-R′, the R′ is usually on the amino    group of the amino acid, and there may be two such substituents. R′    is preferably H; alkyl-CO— (alkyl being up to 25 carbon atoms, such    as up to 17, 19 or 21 carbon atoms and preferably an odd number of    carbon atoms corresponding to a fatty acid carboxylic acid of even    number of carbon atoms or else a low number of carbon atoms such as    1 to 6), especially CH₃—(CH₂)n-CO— where n is for example 1, 2, 4,    12 or 16; alkenyl, especially allyl; haloalkyl-CO—, especially    CF₃—CO—; cycloalkyl-alkyl-CO—, preferably alkyl groupings with a    terminal cyclohexyl group and up to six additional carbon atoms in    the sidechain, especially cyclohexyl-(CH₂)_(n)—CO— where n is for    example 1 or 2; haloalkyl-O—CO—, especially trichloroethoxycarbonyl;    arylalkyl-CO— or arylalkenyl-CO— especially    phenyl-methyl/ethyl/vinyl-CO—, where aryl may be substituted as in    trifluoromethylcinnamoyl; optionally substituted heteroaryl-CO—,    where the substituents and heterocyclic group are as elsewhere    discussed, as in 2-chloronicotinoyl; alkenyl—CO—especially crotonyl;    opitionally subsituted aminoalkyl-CO—, particularly amino acid acyl,    especially alanine, phenylalanine, cysteine, proline, valine,    arginine, tryptophan or other amino acid, or a derivative thereof,    as in Boc-phenylalanine, valine, proline, arginine or tryptophan, or    as in phenethylalanine, trifluoroethylacetylalanine,    trifluorodiacetylalanine and isomers thereof, or diacetyl- or    dipropionyl-trifluoroacetyl, or as in or as in Cbz-Val- or a group    notionally derived from cysteine and being of general formula    Prot^(SH)-S—CH₂—C(═NOProt^(OH))-CO— or    Prot^(SH)-S—CH═C(—OProt^(OH))-CO—, where Prot^(SH) and Prot^(OH) are    protecting groups for thiol and for hydroxy, especially where    Prot^(SH) is Fm and Prot^(OH) is methoxy for the first formulaor MOM    for the second formula; or other possibilities such as a protecting    group as in an alkoxycarbonyl such as Boc, or PhNR′CS. The various    groups may be susbtituted as indicated elsewhere in this    specification.-   R₇ and R₈ are —O—CH₂—O— or R₇ is ═O and R₈ is OMe, especially R₇ and    R₈ are —O—CH₂—O—.-   R₉ is methyl.-   X₂ is —OR″, where R″ is preferably H; alkyl-CO—, especially acetyl;    alkenyl especially allyl; alkenyl-O—CO—, especially allyl-O—CO—;    haloalkyl-CO—, especially trifluoromethylcarbonyl or    chloromethylcarbonyl or 2-chloroethylcarbonyl or    perfluoropropylcarbonyl.

Of special interest are compounds wherein:

-   R₁ is —OR, where R is H or acetyl, alkyl-CO—, especially    n-propyl-CO—, and R₁ is more especially OH.-   R₂ is methoxy.-   R₃ is methyl.-   R₄ is hydrogen.-   R₅ is methyl.-   R₆ is —CN or —OH.-   X₁ is —NHR′, where R′ is preferably alkenyl, especially allyl,    alkyl-CO— (alkyl being 1 to 6 carbon atoms, especially    CH₃—(CH₂)_(n)—CO— where n is for example 1 to 6, and more especially    1 to 4); cycloalkyl-alkyl-CO—, especially cyclohexyl-(CH₂)_(n)—CO    where n is 1 or 2; arylalkyl-CO— or arylalkenyl-CO— especially    phenethylcarbonyl, phenylvinylcarbonyl or benzylcarbonyl,    alkenyl-CO— especially CH₃—CH═CH—CO—; amino acid acyl, especially    Cbz-Val-; optionally substituted heteroaryl-CO—, especially    2-chloropyridinylcarbonyl;-   or X₁ is —NH-aa-R′ where aa is alanine, phenylalanine, tryptophan or    valine; R′ is an amino subsituent and is arylalkyl-CO— especially    phenethylcarbonyl or benzylcarbonyl; alkyl-CO-(alkyl being 1 to 6    carbon atoms, especially CH₃—(CH₂)_(n)—CO— where n is for example 1    to 6 and more especially 1, 2 or 4; alkenyl-CO— especially    CH₃—CH═CH—CO—; or protecting group especially alkyloxy-CO as in Boc;-   or X₁ is —OR′ where R′ is preferably alkyl-CO— (alkyl being 1 to 6    carbon atoms, especially CH₃—(CH₂)n—CO— where n is for example 1 to    6, and more especially 2; arylalkyl-CO— or arylalkenyl-CO—    especially phenethylcarbonyl, phenylvinylcarbonyl or    trifluoromethylcinnamoyl.-   R₇ and R₈ are —O—CH₂—O—.-   R₉ is methyl.-   X₂ is —OR″, where R″ is H; acetyl, allyloxycarbonyl,    chloromethylcarbonyl or perfluoropropylcarbonyl; and R″ is more    especially H; acetyl or allyloxycarbonyl.

Especially preferred embodiments of the present invention are the novelecteinascidin-like compounds with the following general structures I, IIand III that have been prepared from compounds 17, 25, 43 and 45 derivedfrom cyanosafracin B. Compound 25 corresponds to the syntheticintermediate 3 described in U.S. Pat. No. 6,124,292.

Wherein R′, X₂, R₁ and R₆ are each independently selected from thegroups defined below:

R′ X₂ R₁ R₆ H OH OH CN CH₂CH═CH₂ OAc OAc OH COCH₂CH₃ OCH₂CH═CH₂ OMOMCOCH₂CH₂CH₃ OCOOCH₂CH═CH₂ OCOCH₂C₆H₁₁ CO(CH₂)₄CH₃ OCOCF₃ OCOCH₂CH₂C₆H₁₁CO(CH₂)₁₂CH₃ OCOCH₂Cl OCOCH₂CH₂CH₃ CO(CH₂)₁₆CH₃ OCOCH₂CH₂Cl OCO(CH₂)₄CH₃COCH₂C₆H₁₁ OCOCF₂CF₂CF₃ OCO(CH₂)₈CH₃ COCH₂CH₂C₆H₁₁ OCO(CH₂)₁₆CH₃COOCH₂CCl₃ COCH₂Ph COCH₂CH₂Ph COCH═CHCH₃ COCH═CHPh COCH═CHArCF₃COCH(CH₃)NHCOCH₂CH₂Ph CO—(S)—CH(CH₃)NHCOCF₃ CO—(R)—CH(CH₃)NHCOCF₃CO—(S)—CH(NHCbz)CH(CH₃)₂ Boc CSNHPh

In the formulae (XVIIa) or (XVIIb), R¹ is typically aminomethylene,amidomethylene or R¹ with R⁴ forms a group (IV) or (V). Suitableamidomethylene groups include those of formula —CH₂—NH—CO—CHCH₃—NH₂derived from alanine, and similar groups derived from other amino acids,notably, both D and L, glycine, valine, leucine, isoleucine,phenylalanine, tyrosine, tryptophan, methionine, cysteine, aspartate,asparagine, glutamatic acid, glutamine, lysine, arginine, proline,serine, threonine, histidine and hydroxyproline. A general formula forthe group R¹ is then —CH₂—NH-aa, where aa indicates an acyl amino acidgroup.

The group R can be acylated on an —NH, group, and for example N-acylderivatives can be formed from groups —CH₂NH₂ and —CH, —NH-aa. The acylderivatives can be N-acyl or N-thioacyl derivatives thereof, as well ascyclic amides. The acyl groups can illustratively be alkanoyl,haloalkanoyl, arylalkanoyl, alkenoyl, heterocyclylacyl, aroyl,arylaroyl, haloaroyl, nitroaroyl, or other acyl groups. The acyl groupscan be of formula —CO—R^(a), where R^(a) can be various groups such asalkyl, alkoxy, alkylene, arylalkyl, arylalkylene, amino acid acyl, orheterocyclyl, each optionally substituted with halo, cyano, nitro,carboxyalkyl, alkoxy, aryl, aryloxy, heterocyclyl, heterocyclyloxy,alkyl, amino or substituted amino. Other acylating agents includeisothiocyanates, such as aryl isothiocyanates, notably phenylisocyanate. The alkyl, alkoxy or alkylene groups of R^(a) suitably have1 to 6 or 12 carbon atoms, and can be linear, branched or cyclic. Arylgroups are typically phenyl, biphenyl or naphthyl. Heterocyclyl groupscan be aromatic or partially or completely unsaturated and suitably have4 to 8 ring atoms, more preferably 5 or 6 ring atoms, with one or moreheteroatoms selected from nitrogen, sulphur and oxygen.

Without being exhaustive, typical R^(a) groups include alkyl, haloalkyl,alkoxyalkyl, haloalkoxyalkyl, arylalkylene, haloalkylarylakylene, acyl,haloacyl, arlyalkyl, alkenyl and amino acid. For example, R^(a)—CO— canbe acetyl, trifluoroacetyl, 2,2,2-trichloroethoxycarbonyl,isovalerylcarbonyl, trans-3-(trifluoromethyl)cinnamoylcarbonyl,heptafluorobutyrylcarbonyl, decanoylcarbonyl, trans-cinnamoylcarbonyl,butyrylcarbonyl, 3-chloropropyonylcarbonyl, cinnamoylcarbonyl,4-methylcinnamoylcarbonyl, hydrocinnamoylcarbonyl, ortrans-hexenoylcarbonyl, or alanyl, arginyl, aspartyl, asparagyl, cystyl,glutamyl, glutaminyl, glycyl, histidyl, hydroxyprolyl., isoleucyl,leucyl, lysyl, methionyl, phenylalanyl, prolyl, seryl, threonyl,thyronyl, tryptophyl, tyrosyl, valyl, as well as other less common aminoacid acyl groups, as well as phthalimido and other cyclic amides. Otherexamples may be found among the listed protecting groups.

Compounds wherein —CO—R^(a) is derived from an amino acid and include anamino group can themselves form acyl derivatives. Suitable N-acylcommands include dipeptides which in turn can form N-acyl derivatives.

In an important aspect of this invetnion, there are provided preferredcompounds of the formula:

wherein:

-   R¹ is —CH₂—N(R^(a))₂ or CH₂—OR^(a), where R^(a) is H; alkyl-CO—;    haloalkyl-CO—; cycloalkylalkyl-CO—; haloalkyl-O—CO—; arylalkyl-CO—;    arylalkenyl-CO—; heteroaryl-CO—; alkenyl-CO—; alkenyl; amino acid    acyl; or a protecting group;-   R⁵ is —OR″, where R″ is H; alkyl-CO—; cycloalkyl-CO—; haloalkyl-CO—    or a protecting group;-   R¹⁸ is —OR, where R is H, alkyl-CO—; cycloalkylalkyl-CO—; or a    protecting group;-   R²¹ is —CN or —OH.

Typically such a compound is of the formula:

wherein R¹, R⁵, R¹⁸, and R²¹ are as defined.

In such preferred compounds of this invention, R¹ can be —CH₂—NHR^(a).

R^(a) can be -aa-R^(b) where aa is amino acid acyl and R^(b) is asdefined for R^(a). The amino acid acyl is optionally further substitutedwith one or more R^(a) groups.

In further preferred comopunds, R¹ is CH₂—NH-aa-R^(b) where aa is anamino acid and R^(b) is hydrogen; protecting group; arylalkenyl-CO—;haloalkyl-CO—; alkyl-CO—, arylalkyl-CO—, or amino acid acyl. Suchcomopunds include those wherein R¹ is —CH₂—NH-aa-R^(b) where aa isalanine and R^(b) is hydrogen, Boc, PhNHCS—, CF₃CO—, PhNAcCS—,trifluorocinnamoyl cinnamoyl, C₃F₇CO—, butyryl, 3-chloroproprionoyl,hydrocinnamoyl, hexanoyl, phenylacetyl, Cbz-val or acetyl; —CH₂-aa-R^(b)where aa is valine and R^(b) is Cbz or Boc; —CH₂-aa-R^(b) where aa isphenylalanine and R^(b) is Boc; —CH₂-aa-R^(b) where aa is proline andR^(b) is Boc: —CH₂-aa-R^(b) where aa is arginine and R^(b) is Boc; orCH₂-aa-R^(b) where aa is tryptophan and R^(b) is Boc.

R¹ can be —CH₂—NR^(a)-aa-R^(b) where aa is an amino acid, R^(a) isalkyl-CO— and R^(b) is haloalkyl-CO—. Such compounds include thosewherein R¹ is —CH₂—NR^(a)-aa-R^(b) where aa is acetylalanine, R^(a) isacetyl or butyryl, and R^(b) is CF₃—CO—.

R¹ can be —CH₂—NHR^(a) where R^(a) is hydrogen, protecting group,alkyl-CO—; alkenyl-CO—; arylalkenyl-CO—; arylalkyl-CO—; heteroaryl-CO—;cycloalkylalkyl-CO—; or alkenyl. Such compounds include those wherein R¹is —CH₂—NHR^(a) where R^(a) is hydrogen, Troc, acetyl; isovaleroyl,decanoyl, cinnamoyl, hydrocinnamoyl, phenylacetyl, propionyl myristoyl,stearoyl, hexanoyl, crotonyl, chloronicotinoyl, cyclohexylacetyl,cyclohexylpropionyl or allyl.

R¹ can be —CH₂—OR^(a) where R^(a) is hydrogen; a protected cysteine; acysteine derivative of the formula Prot^(SH)-S—CH₂—C(NHProt^(NH))-CO—,where Prot^(SH) and Prot^(NH) are protecting groups for thiol and foramino; a protecting group; alkyl-CO—; arylalkyl-CO—; arylalkenyl-CO—;

a cysteine derivative of the formula Prot^(SH)-S—CH₂—C(═NOProt^(OH))-CO—where Prot^(SH) and Prot^(OH) are protecting groups for thiol and forhydroxy; or a cysteine derivative of formulaProt^(SH)-S—CH═C(—OProt^(OH))-CO—, where Prot^(SH) and Prot^(OH) areprotecting groups for thiol and for hydroxy. Such compounds includethose wherein R¹ is —CH₂—OR^(a) where R^(a) is hydrogen;S—Fm—O-TBDMS-cysteine; a cysteine derivative of the formulaProt^(SH)-S—CH₂—C(NHProt^(NH))-CO—, where Prot^(SH) is Fm and Prot^(OH)is Troc; TBDPS; butyryl; trfiluormethylcinnamoyl; cinnamoyl;hydrocinnamoyl; a cysteine derivative of the formulaProt^(SH)-S—CH₂—C(═NOProt^(OR))-CO— where Prot^(SH) is Fm and Prot^(OH)is methoxy; or a cysteine derivative of formula Prot where Prot^(SH) isFm and Prot^(OH) is MOM.

In these preferred compounds, R⁵ is suitably —OR″, where R″ is H;alkyl-CO where the alkyl has an odd number of carbon atoms,ω-cyclohexylalkyl-CO—; or a protecting group.

In these preferred compounds, R¹⁸ is suitably —OR, where R is H,alkyl-CO—; or a protecting group;

In one variation which relates to intermediate products, the ring A ismodified to incorporate the substructure shown as formula (XX) or (XXI),discussed later.

In another variation relating to intermediates, the group R¹ can be—CH₂O—CO—CFu-CH₂—S-Prot³, derived from a compound of formula (XIX),where Prot³ and Fu have the indicated meanings. In such a case, R⁷ andR⁸ from the oxymethyleneoxy group. The group R¹⁸ is usually protected.Usually R²¹ is cyano.

Preferably R^(14a) and R^(14b) are hydrogen. Preferably R¹⁵ is hydrogen.The O-acyl derivatives are suitably aliphatic O-acyl derivatives,especially acyl derivatives of 1 to 4 carbon atoms, and typically anO-acetyl group, notably at the 5-position.

Suitable protecting groups for phenols and hydroxy groups include ethersand esters, such as alkyl, alkoxyalkyl, aryloxyalkyl, alkoxyalkoxyalkyl,alkylsilylalkoxyalkyl, alkylthioalkyl, arylthioalkyl, azidoalkyl,cyanoalkyl, chloroalkyl, heterocyclic, arylacyl, haloarylacyl,cycloalkylalkyl, alkenyl, cycloalkyl, alyklarylalkyl, alkoxyarylalkyl,nitroarylalkyl, haloarylalkyl, alkylaminocarbonylarylalkyl,alkylsulfinylarylalky, alkylsilyl and other ethers, and arylacyl, arylalkyl carbonate, aliphatic carbonate, alkylsulfinylarlyalkyl carbonate,alkyl carbonate, aryl haloalkyl carbonate, aryl alkenyl carbonate, arylcarbamate, alkyl phosphinyl, alkylphosphinothioyl, aryl phosphinothioyl,aryl alkyl sulphonate and other esters. Such groups may optionally besubstituted with the previously mentioned groups in R¹.

Suitable protecting groups for amines include carbamates, amides, andother protecting groups, such as alkyl, arylalkyl, sulpho- orhalo-arylalkyl, haloalkyl, alkylsilylalkyl, arylalkyl, cycloalkylalkyl,alkylarylalkyl, heterocyclylalkyl nitroarylalkyl acylaminoalkyl,nitroaryldithioarylalkyl, dicycloalkylcarboxamidoalkyl, cycloalkyl,alkenyl, arylalkenyl, nitroarylalkenyl, heterocyclylalkenyl,heterocyclyl, hydroxyheterocyclyl, alkyldithio, alkoxy- or halo- oralkylsulphinyl arylalkyl, hetercyclylacyl, and other carbamates, andalkanoyl, haloalkanoyl, arylalkanoyl, alkenoyl, heterocyclylacyl, aroyl,arylaroyl, haloaroyl, nitroaroyl, and other amides, as well as alkyl,alkenyl, alkylsilylalkoxyalkyl, alkoxyalkyl, cyanoalkyl, heterocyclyl,alkoxyarylalkyl, cycloalkyl, nitroaryl, arylalkyl, alkoxy- orhydroxy-arylalkyl, and many other groups. Such groups may optionally besubstituted with the previously mentioned groups in R¹.

Examples of such protecting groups are given in the following tables,protection for —OH group

abbreviation ethers methyl methoxymethyl MOM benzyloxymethyl BOMmethoxyethoxymethyl MEM 2-(trimethylsilyl)ethoxymethyl SEMmethylthiomethyl MTM phenylthiomethyl PTM azidomethyl cyanomethyl2,2-dichloro-1,1-difluoroethyl 2-chloroethyl 2-bromoethyltetrahydropyranyl THP 1-ethoxyethyl EE phenacyl 4-bromophenacylcyclopropylmethyl allyl propargyl isopropyl cyclohexyl t-butyl benzyl2,6-dimethylbenzyl 4-methoxybenzyl MPM or PMB o-nitrobenzyl2,6-dichlorobenzyl 3,4-dichlorobenzyl 4-(dimethylamino)carbonylbenzyl4-methylsuflinylbenzyl Msib 9-anthrylmethyl 4-picolylheptafluoro-p-tolyl tetrafluoro-4-pyridyl trimethylsilyl TMSt-butyldimethylsilyl TBDMS t-butyldiphenylsilyl TBDPS triisopropylsilylTIPS esters aryl formate aryl acetate aryl levulinate aryl pivaloateArOPv aryl benzoate aryl 9-fluorocarboxylate aryl methyl carbonate1-adamantyl carbonate t-butyl carbonate BOC-OAr 4-methylsulfinylbenzylcarbonate Msz-Oar 2,4-dimethylpent-3-yl carbonate Doc-Oar aryl2,2,2-trichloroethyl carbonate aryl vinyl carbonate aryl benzylcarbonate aryl carbamate dimethylphosphinyl Dmp-OArdimethylphosphinothioyl Mpt-OAr diphenylphosphinothioyl Dpt-Oar arylmethanesulfonate aryl toluenesulfonate aryl 2-formylbenzenesulfonateprotection for the —NH₂ group carbamates methyl ethyl 9-fluorenylmethylFmoc 9-(2-sulfo)fluroenylmethyl 9-(2,7-dibromo)fluorenylmethyl17-tetrabenzo[a,c,g,i]fluorenylmethyl Tbfmoc 2-chloro-3-indenylmethylClimoc benz[f]inden-3-ylmethyl Bimoc2,7-di-t-butyl[9-(10,10-dioxo-10,10,10,10- DBD-Tmoctetrahydrothioxanthyl)]methyl 2,2,2-trichloroethyl Troc2-trimethylsilylethyl Teoc 2-phenylethyl hZ1-(1-adamantyl)-1-methylethyl Adpoc 2-chlooethyl1,1-dimethyl-2-chloroethyl 1,1-dimethyl-2-bromoethyl1,1-dimethyl-2,2-dibromoethyl DB-t-BOC 1,1-dimethyl-2,2,2-trichloroethylTCBOC 1-methyl-1-(4-biphenyl)ethyl Bpoc1-(3,5-di-t-butylphenyl)-1-1-methylethyl t-Burmeoc 2-(2′-and4′-pyridyl)ethyl Pyoc 2,2-bis(4′-nitrophenyl)ethyl Bnpeocn-(2-pivaloylamino)-1,1-dimethylethyl2-[(2-nitrophenyl)dithio]-1-phenylethyl NpSSPeoc2-(n,n-dicyclohexylcarboxamido)ethyl t-butyl BOC 1-adamantyl 1-Adoc2-adamantyl 2-Adoc vinyl Voc allyl Aloc or Alloc 1-isopropylallyl Ipaoccinnamyl Coc 4-nitrocinnamyl Noc 3-(3′-pyridyl)prop-2-enyl Paloc8-quinolyl n-hydroxypiperidinyl alkyldithio benzyl Cbz or Zp-methoxybenzyl Moz p-nitrobenzyl PNZ p-bromobenzyl p-chlorobenzyl2,4-dichlorobenzyl 4-methylsulfinylbenzyl Msz 9-anthrylmethyldiphenylmethyl phenothiazinyl-(10)-carbonyln′-p-toluenesulfonylaminocarbonyl n′-phenylaminothiocarbonyl amidesformamide acetamide chloroacetamide trifluoroacetamide TFAphenylacetamide 3-phenylpropanamide pent-4-enamide picolinamide3-pyridylcarboxamide benzamide p-phenylbenzamide n-phthalimiden-tetrachlorophthalimide TCP 4-nitro-n-phthalimide n-dithiasuccinimideDts n-2,3-diphenylmaleimide n-2,5-dimethylpyrrolen-2,5-bis(triisopropylsiloxyl)pyrrole BIPSOPn-1,1,4,4-tetramethyldisiliazacyclopentante adduct STABASE1,1,3,3-tetramethyl-1,3-disilaisoindoline BSB special —NH protectivegroups n-methylamine n-t-butylamine n-allylaminen-[2-trimethylsilyl)ethoxy]methylamine SEM n-3-acetoxypropylaminen-cyanomethylamine n-(1-isopropyl-4-nitro-2-oxo-3-pyrrolin-3-yl)aminen-2,4-dimethoxybenzylamine Dmb 2-azanorbornenes n-2,4-dinitrophenylaminen-benzylamine Bn n-4-methoxybenzylamine MPM n-2,4-dimethoxybenzylamineDMPM n-2-hydroxybenzylamine Hbn n-(diphenylmethyl)amino DPMn-bis(4-methoxyphenyl)methylamine n-5-dibenzosuberylamine DBSn-triphenylmethylamine Tr n-[(4-methoxyphenyl)diphenylmethyl]amino MMTrn-9-phenylflurenylamine Pf n-ferrocenylmethylamine Fcm n-2-picolylaminen′-oxide n-1,1-dimethylthiomethyleneamine n-benzylideneaminen-p-methoxybenzylideneamine n-diphenylmethyleneaminen-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine n-nitroamine n-nitrosoaminediphenylphosphinamide Dpp dimethylthiophosphinamide Mptdiphenylthiophosphinamide Ppt dibenzyl phosphoramidate2-nitrobenzenesulfenamide Npsn-1-(2,2,2-trifluoro-1,1-diphenyl)ethylsufenamide TDE3-nitro-2-pyridinesulfenamide Npys p-toluenesulfonamide Tsbenzenesulfonamide

Examples of preferred methods of this invention will firstly beconsidered with reference to starting compunds 45, 43 and 25. It will beappreicated that the particular substituents, notably at positions C-5and C-18, can be varied in the light of the present disclosure.

The preferred methods of producing the compounds of formula I, II andIII are described below in the following reaction schemes with examplesof typical substituent groups.

As illustrated in Scheme 1 the first step for producing the preferredcompounds (I) (where R₁=OH, X₂=OAc and R₆=CN or OH) of the presentinvention from compound 45 is the high yielding conversion of the aminogroup to the amide group.

After acylation of the amino group the second step is the transformationof the CN group into an OH group by reaction with silver nitrate inAcCN/H₂O.

The preparation of other compounds of the general formula I of thepresent invention starting from compound 17 is described below (Scheme4).

As illustrated in Scheme 2 another group of interesting derivatives withformula II (where R₁═OH, X₂═OAc and R₆═CN or OH) can be obtained fromcompound 43 using the following sequence. Acylation of the amino groupto provide the corresponding amide and transformation of the CN groupinto an OH group by reaction with silver nitrate in AcCN/H2O.

The preparation of other compounds of the general formula II of thepresent invention starting from compound 17 is described below (Scheme4).

The preferred procedure for producing compounds of formula III is thetransformation of compound 25 into the corresponding ester derivativesby acylation of the OH group, deprotection of the phenol group followedby acetylation and deprotection of the MOM group to provide thecorresponding ester followed by transformation of the CN group to the OHgroup by reaction with silver nitrate in AcCN/H₂O to give the compoundof formula III (where R₁═OH, X₂═OAc and R₆═CN or OH).

Other compounds of the general formulae I and II of the presentinvention can be prepared from compound 17 via the amine intermediate120 as described in Scheme 4.

The following additional compounds of the present invention (includingfor example 140 and 141) have been prepared starting from cyanosafracinB (2) as described in detail in the examples (Scheme 5).

As the skilled artisan will readily appreciate the reaction schemesdescribed herein may be modified and/or combined in various ways, andthe compounds generated therefore are to be considered as being part ofthis invention. In particular the starting material and/or reagents andreactions can be varied to suit other combinations of the substituentgroups in the formulae I, II and III.

In a related aspect, the present invention is directed at the use of aknown compound, safracin B, also referred to as quinonamine, inhemisynthetic synthesis.

More generally, the invention relates to a hemisynthetic process for theformation of intermediates, derivatives and related structures ofecteinascidin or other tetrahydroisoquinolinephenol compounds startingfrom natural bis(tetrahydroisoquinoline) alkaloids. Suitable startingmaterials for the hemi-synthetic process include the classes ofsaframycin and safracin antibiotics available from different culturebroths, and also the classes of reineramicin and xestomycin compoundsavailable from marine sponges.

A general formula (XV) for the starting compounds is as follows:

where:

-   R¹ is an amidomethylene group such as    —CH₂—NH—CO—CR^(25a)R^(25b)R^(25c) where R^(25a) and R^(25b) form a    keto group or one is —OH, —NH₂ or —OCOCH₃ and the other is    —CH₂COCH₃, —H, —OH or —OCOCH₃, provided that when R^(25a) is —OH or    —NH₂ then R^(25b) is not —OH, and R^(25c) is —H, —CH₃ or —CH₂CH₃, or    R¹ is an acyloxymethylene group such as —CH, —O—CO—R, where R is    —C(CH₃)═CH—CH₃ or —CH₃;-   R⁵ and R⁸ are independently chosen from —H, —OH or —OCOCH₂OH, or R⁵    and R⁸ are both keto and the ring A is a p-benzoquinone ring;-   R^(14a) and R^(14b) are both —H or one is —H and the other is —OH,    —OCH₃ or —OCH₂CH₃, or R^(14a) and R^(14b) together form a keto    group,-   R¹⁵ and R¹⁸ are independently chosen from —H or —OH, or R⁵ and R⁸    are both keto and the ring A is a p-benzoquinone ring; and-   R²¹ is —OH or —CN.

A more general formula for these class of compounds is provided below:

-   wherein the substituent groups defined by R₁, R₂, R₃, R₄, R₅, R₆,    R₇, R₈, R₉, R₁₀ are each independently selected from the group    consisting of H, OH, OCH₃, CN, ═O, CH₃;-   wherein X are the different amide or ester functionalities contained    in the mentioned natural products;-   wherein each dotted circle represents one, two or three optional    double bonds.

Thus, according to the present invention, we now provide hemisyntheticroutes for the production of intermediates including Intermediate 11 andthus for the production of the ecteinascidin compounds as well asphthalascidin and additional compounds. The hemisynthetic routes of theinvention each comprise a number of transformation steps to arrive atthe desired product. Each step in itself is a process in accordance withthis invention. The invention is not limited to the routes that areexemplified, and alternative routes may be provided by, for example,changing the order of the transformation steps, as appropriate.

In particular, this invention involves the provision of a 21-cyanostarting material of general formula (XVI):

where R¹, R⁵, R⁸, R^(14a), R^(14b), R¹⁵ and R¹⁸ are as defined.

Other compounds of formula (XVI) with different substituents at the21-position may also represent possible starting materials. In general,any derivative capable of production by nucleophilic displacement of the21-hydroxy group of compounds of formula (XV) wherein R²¹ is a hydroxygroup cis a candidate. Examples of suitable 21-substituents include butare not limited to:

-   a mercapto group;-   an alkylthio group (the alkyl group having from 1 to 6 carbon    atoms);-   an arylthio group (the aryl group having from 6 to 10 carbon atoms    and being unsubstituted or substituted by from 1 to 5 substituents    selected from, for example, alkyl group having from 1 to 6 carbon    atoms, alkoxy groups having from 1 to 6 carbon atoms, halogen atoms,    mercapto groups and nitro groups);-   an amino group;-   a mono-or dialkylamino (the or each alkyl group having from 1 to 6    carbon atoms);-   a mono-or diarylamino group (the or each aryl group being as defined    above in relation to arylthio groups);-   an α-carbonylalkyl group of formula —C(R^(a))(R^(b))—C(═O)R^(c),    where R^(a) and R^(b) are selected from hydrogen atoms, alkyl groups    having from 1 to 20 carbon atoms, aryl groups (as defined above in    relation to arylthio groups) and aralkyl groups (in which an alkyl    group having from 1 to 4 carbon atoms is substituted by an aryl    group a defined above in relation to arylthio groups), with the    proviso that one of R^(a) and R^(b) is a hydrogen atom;-   R^(c) is selected from a hydrogen atom, an alkyl group having from 1    to 20 carbon atoms, aryl groups (as defined above in relation to    arylthio groups), an aralkyl group (in which an alkyl group having    from 1 to 4 carbon atoms is substituted by an aryl group a defined    above in relation to arylthio groups), an alkoxy group having from 1    to 6 carbon atoms, an amino group or a mono- or dialkylamino group    as defined above.

Thus, in a more general aspect, the present invention relates toprocesses where the first step is to form a 21-deriviative using anucleophilic reagent. We refer to such compounds as 21-Nuc compounds.

The presence of the 21-cyano group is required for some of theend-products, notably ecteinascidin 770 and phthalascidin, while forother end-products it acts as a protecting group which can readily beconverted to another substituent, such as the 21-hydroxy group ofecteinascidin 743 or of 21-hydroxyphthalascidin. The adoption of the21-cyano compound as the starting material effectively stabilises themolecule during the ensuing synthetic steps, until it is optionallyremoved. Other 21-Nuc compounds can offer this and other advantages.

In one important aspect, the present invention consists in the use of a21-cyano compound of the general formula (XVI) in the preparation of abis- or tris-tetrahydroisoquinolinephenol) compounds. Products which maybe prepared include intermediates such as Intermediate 11, and theecteinascidins and phthalascidin, as well as new and known compounds ofrelated structure.

Preferred starting materials include those compounds of formula (XV) or(XVI) where R^(14a) and R^(14b) are both hydrogen. Preferred startingmaterials also include compounds of formula (XV) or (XVI) where R¹⁵ ishydrogen. Furthermore, the preferred starting materials includecompounds of formula (XV) or (XVI) where ring E is a phenolic ring.Preferred starting materials further include compounds of formula (XV)or (XVI) where at least one, better at least two or three of R⁵, R⁸, R¹⁵and R¹⁸ is not hydrogen.

Examples of suitable starting materials for this invention includesaframycin A, saframycin B, saframycin C, saframycin G, saframycin H,saframycin S, saframycin Y₃, saframycin Yd₁, saframycin Ad₁, saframycinYd₂, saframycin AH₂, saframycin AH₂Ac, saframycin AH₁, saframycin AH₁Ac,saframycin AR₃, renieramycin A, renieramycin B, renieramycin C,renieramycin D, renieramycin E, renieramycin F, xestomycin, saframycinD, saframycin F, saframycin Mx-1, saframycin Mx-2, safracin A, safracinB and saframycin R, Preferred starting materials have a cyano group inposition 21, for the group R²¹.

In a particularly preferred aspect, the invention involves ahemisynthetic process wherein the transformation steps are applied tosafracin B:

Safracin B presents a ring system closely related to the ecteinascidins.This compound has the same pentacycle structure and the samesubstitution pattern in the right-band aromatic ring, ring E. Also,safracin B presents very close similarities to some of the syntheticintermediates in the total synthesis of ET-743, particularly to theintermediate 11. Such intermediate can be transformed into Et-743 usinga well established method. Synthetic conversion of safracin B intointermediate 11 will therefore provide an hemi-synthetic method toobtain ET-743.

Thus, we provide Intermediate 11 made from this compound safracin B, andcompounds derived from Intermediate 11, particularly ecteinascidincompounds. We further provide phthalascidin made from safracin B. Theinvention also relates to use of safracin B in the production ofIntermediate 11, phthalascidin, ecteinascidin compounds and the otherintermediates of the invention. The invention also relates to compoundsdescribed herein derived from the other suggested starting materials,and use of those compounds in the production of such compounds.

The more preferred starting materials of this invention have a 21-cyanogroup. The currently most preferred compound of the present invention isthe compound of Formula 2. This compound is obtained directly fromsafracin B and is considered a key intermediate in the hemisyntheticprocess.

In a related aspect, we provide cyanosafracin B by fermentation of asafracin B-producing strain of Pseudomonas fluorescens, and working upthe cultured broth using cyanide ion. The preferred strain ofPseudomonas fluorescens is strain A2-2, FERM BP-14, which is employed inthe procedure of EP 055,299. A suitable source of cyanide ion ispotassium cyanide. In a typical work-up, the broth is filtered andexcess cyanide ion is added. After an appropriate interval of agitation,such as 1 hour, the pH is rendered alkaline, say pH 9.5, and an organicextraction gives a crude extract which can be further purified to givethe cyanosafracin B.

Safracin B includes an alanyl sidechain. In one aspect of the invention,we have found that protection of the free amino group with a Boc groupcan give strong advantages.

In general, the conversion of the 21-cyano starting compound to anecteinascidin analog of this invention can be carried out in accordancewith our copending PCT patent application, attorney reference wpp83894,which also claims priority from the PCT filing published as WO 0069862published 23 Nov. 2000, and which relates to hemisynthetic methods andnew compounds. We incorporate the text of the copending PCT application,attorney reference wpp83894, by reference to the extent that there isdisclosure therein which is not in the present specification.

Typically the hemisynthesis of an analog of this invention involves:

-   a) conversion if necessary of a quinone system for the ring E into    the phenol system-   b) conversion if necessary of a quinone system for the ring A into    the phenol system;-   c) conversion of the phenol system for the ring A into the    methylenedioxyphenol ring; and-   d) derivatisation as appropriate, such as acylation.

Step (a), conversion if necessary of a quinone system for the ring Einto the phenol system, can be effected by conventional reductionprocedures. A suitable reagent system is hydrogen with apalladium-carbon catalyst, though other reducing systems can beemployed.

Step (b), conversion if necessary of a quinone system for the ring Ainto the phenol system is analogous to step (a), and more detail is notneeded.

Step (c), conversion of the phenol system for the ring A into themethylenedioxyphenol ring, can be effected in several ways, possiblyalong with step (b). For example, a quinone ring A can be demethylatedin the methoxy substituent at the 7-position and reduced to adihydroquinone and trapped with a suitable electrophilic reagent such asCH₂Br₂, BrCH₂Cl, or a similar divalent reagent directly yielding themethylenedioxy ring system, or with a divalent reagent such asthiocarbonyldiimidazol which yields a substituted methylenedioxy ringsystem which can be converted to the desired ring.

Derivatisation in step (d) can include acylation, for instance with agroup R^(a)—CO— as well as conversion of the 12-NCH₃ group to 12-NH or12-NCH₂CH₃. Such conversion can be effected before or after the othersteps, using available methods.

By way of illustration, it is now feasible to transform cyanosafracin Bin a shorter and more straightforward way to make new analogs.Cyanosafracin B can be transformed into Intermediate 25;

and from this derivative it is possible to introduce further analogs ofthis invention.

One method of this invention transforms cyanosafracin B intointermediate 25 through a sequence of reactions that involvesessentially (1) removal of methoxy group placed in ring A, (2) reductionof ring A and formation of methylene-dioxy group in one pot, (3)hydrolysis of amide function placed over carbon 1, (4) transformation ofthe resulting amine group into hydroxyl group.

The conversion of the 2-cyano compound into Intermediate 25 usuallyinvolves the following steps (see scheme II):

-   formation of the protected compound of Formula 14 by reacting 2 with    tert-butoxycarbonyl anhydride;-   converting of 14 into the di-protected compound of Formula 15 by    reacting with bromomethylmethyl ether and diisopropylethylamine in    acetonitrile;-   selectively elimination of the methoxy group of the quinone system    in 15 to obtain the compound of Formula 16 by reacting with a    methanolic solution of sodium hydroxide;-   transforming of 16 into the methylene-dioxy compound of Formula 18    by employing the next preferred sequence: (1) quinone group of    compound 16 is reduced with 10% Pd/C under hydrogen atmosphere; (2)    the hydroquinone intermediate is converted into the methylenedioxy    compound of Formula 17 by reacting with bromochloromethane and    caesium carbonate under hydrogen atmosphere; (3) 17 is transformed    into the compound of Formula 18 by protecting the free hydroxyl    group as a OCH₂R group. This reaction is carried out with BrCH₂R and    caesium carbonate, where R can be aryl, CH═CH₂ OR′ etc.-   elimination of the tert-butoxycarbonyl and the methyloxymethyl    protecting groups of 18 to afford the compound of Formula 19 by    reacting with a solution of HCl in dioxane. Also this reaction is    achieved by mixing 18 with a solution of trifluoroacetic acid in    dichloromethane;-   formation of the thiourea compound of Formula 20 by reacting 19 with    phenylisothiocyanate;-   converting compound of Formula 20 into the amine compound of Formula    21 by reacting with a solution of hydrogen chloride in dioxane;-   transforming compound of Formula 21 into the N-Troc derivative 22 by    reacting with trichloroethyl chloroformate and pyridine;-   formation of the protected hydroxy compound of Formula 23 by    reacting 22 with bromomethylmethyl ether and diisopropylethylamine;-   transforming compound of Formula 23 into the N—H derivative 24 by    reacting with acetic acid and zinc;-   conversion of compound of Formula 24 into the hydroxy compound of    Formula 25 by reaction with sodium nitrite in acetic acid.    Alternatively, it can be used nitrogen tetroxide in a mixture of    acetic acid and acetonitrile followed by treatment with sodium    hydroxide. Also, it can be used sodium nitrite in a mixture of    acetic anhydride-acetic acid, followed by treatment with sodium    hydroxide.

The conversion of the Intermediate 25 compound into other analogs ofthis invention is then readily achieved, as illustrated for example inScheme III, which usually involves the following steps:

-   transforming compound of formula 24 into the derivative 30 by    protecting the primary hydroxyl function with    (S)—N-2,2,2-tricloroethoxycarbonyl-S-(9H-fluoren-9-ylmethyl)cysteine    29;-   converting the protected compound of formula 30 into the phenol    derivative 31 by cleavage of the allyl group with tributyltin    hydride and dichloropalladium-bis (triphenylphosphine), transforming    the phenol compound of Formula 31 into compound of formula 32 by    oxidation with benzeneseleninic anhydride at low temperature;

The route described above to transform Intermediate 25 can beconveniently modified to form other derivatives.

In more detail, the conversion of the starting 21-cyano compound to arelated product of this invention, such as one of formula (XX), usuallyinvolves the following steps:

-   a) conversion if necessary of a quinone system for the ring E into    the phenol system-   b) formation of the —R⁵ group at the 5-position in ring A;-   c) formation of the R¹ group at the 1-position in ring B; and-   d) conversion if necessary of a quinone system for the ring A into    the phenol system;-   e) conversion of the phenol system for the ring A into the    methylenedioxyphenol ring.

These steps have many similarities with the steps given previously. Step(c) typically involves forming a group —CH₂NH₂ at the 1-position andacylating it.

Phthlascidin can be made using Intermediates described in the conversionof cyanosafracin B into Intermediate 25. For example, Intermediates 21and 17 are suitable starting materials to make Phthlascidin and otheranalogs of this invention.

As shown in scheme V, the process for the synthetic formation ofphthlascidin starting from Intermediate 21 comprises the sequentialsteps of:

-   transforming of 21 into the compound of Formula 27 by reaction with    phthalic anhydride in dichloromethane and carbonyldiimidazole.-   converting of 27 into phthlascidin by reacting with tributyltin    hydride and dichloro palladium-bis(triphenylphosphine) or basic    media, followed by reaction with acetyl chloride.

As shown in scheme VI, the process for the synthetic formation ofphthlascidin starting from Intermediate 17 comprises the sequentialsteps of:

-   acetylation of the hydroxyl group of compound of formula 17 with    acetyl chloride and pyridine to give the acetylated intermediate    compound of formula 42;-   removal of the tert-butoxycarbonyl and the methyloxymethyl    protecting groups of 42 to afford the compound of Formula 43 by    reacting with a solution of HCl in dioxane. Also this reaction is    achieved by mixing 42 with a solution of trifluoroacetic acid in    dichloromethane;-   formation of the thiourea compound of Formula 44 by reacting 43 with    phenylisothiocyanate;-   converting compound of Formula 44 into the amine compound of Formula    45 by reacting with a solution of hydrogen chloride in dioxane;-   transforming of 45 into phthlascidin by reaction with phthalic    anhydride in dichloromethane and carbonyldiimidazole.    Other analogs can be made for example from 43 or 45 by a similar    manner.

The conversion of the 21-cyano compound to Intermediate 11 or a relatedintermediate of formula (XXI) usually involves the following steps:

-   a) conversion if necessary of a quinone system for the ring E into    the phenol system-   b) formation of the —OProt¹ group at the 18-position, in ring E;-   c) formation of the —CH₂—OProt² group at the 1-position, in ring B;    and-   d) conversion if necessary of a quinone system for the ring A into    the phenol system;-   e) conversion of the phenol system for the ring A into the    methylenedioxyphenol ring.

Step (b), formation of the —OProt¹ group at the 18-position in ring E,is a typical protection reaction for a phenol group, and no specialcomments need to be made. Appropriate conditions are chosen depending onthe nature of the protecting group. The other steps are similar to theother reactions.

Step (b), formation of the —CH₂—OProt² group at the 1-position in ringB, is normally carried out by forming a group —CH₂NH₂ at the 1-positionand then converting the amine function to a hydroxy function andprotecting. Thus, where the starting material has a group R¹ which is—CH₂—NH—CO—CR^(25a)R^(21b)R^(25c) then it is matter of removing theN-acyl group. Where the starting material has a group R¹ which is—CH₂—O—CO—R¹ then no change may be needed for an ecteinascidin productwhere the substituent R¹ is the same. For other products, it is matterof removing the O-acyl group. Various procedures are available for suchde-acylations. In one variation, the deacylation and conversion to ahydroxy function are performed in one step. Thereafter, the hydroxygroup can be acylated or otherwise converted to give the appropriate R¹group.

U.S. Pat. No. 5,721,362 describe synthetic methods to make ET-743through a long multistep synthesis. One of the Intermediates of thissynthesis is Intermediate 11. Using cyanosafracin B as starting materialit is possible to reach Intermediate 11 providing a much shorter way tomake such Intermediate and therefor improving the method to make ET-743Cyanosafracin B can be converted into Intermediate 25 by the methodsdescribed above. From Intermediate 25 is possible to reach Intermediate11 using the following steps, see scheme VII.

formation of the protected hydroxy compound of Formula 26 by reacting 25with tert-butyldiphenylsilyl chloride in the presence of a base;

final cleavage of the allyl group with tributyltin hydride anddichloropalladium-bis (triphenylphosphine) in 26 that leads to theformation of the intermediate 11.

One embodiment of the synthetic process of the present invention totransform safracin B into intermediate 11 is a modification andextension of Scheme VIII and comprises the sequential steps of:

-   stereospecifically converting the compound Safracin B to the    compound of Formula 2 by selective replacement of OH by CN by    reacting with KCN in acid media,-   forming the thiourea compound of Formula 3 by reacting compound of    Formula 2 with phenyl isothiocyanate;-   converting the thiourea compound of Formula 3 into the acetamide of    Formula 5 by an hydrolysis in acid media followed by addition of    acetic anhydride; The intermediate amine compound of Formula 4 can    be isolated by quenching the hydrolysis in acid media with sodium    bicarbonate, but this intermediate is highly unstable, and is    transformed quickly into a five member cyclic imine, named compound    6;-   forming the protected compound of Formula 7 by reacting with    bromomethylmethyl ether and diisopropylethylamine in    dichloromethane;-   selectively de-methylating the methoxy group of the quinone system    of compound of Formula 7 into the compound of Formula 8 by reacting    with methanolic solution of sodium hydroxide;-   transforming the compound of Formula 8 into methylenedioxy-compound    of Formula 9 by the preferred following sequence: (1) quinone group    of compound 8 is reduced with 10% Pd/C under hydrogen    atmosphere; (2) the hydroquinone intermediate is converted into the    methylene-dioxy compound of Formula 9 by reacting with    bromochloromethane and cesium carbonate under hydrogen    atmosphere; (3) compound of Formula 9 is transformed into compound    of Formula 10 by protecting the free hydroxyl group as a OCH₂R    group, by reacting with BrCH₂R and cesium carbonate, where R can be    aryl, CH═CH₂, OR′ etc.; converting the acetamide group of compound    of Formula 10 into the corresponding hydroxyl group of Formula 11 by    reaction with nitrogen tetroxide in a mixture of acetic acid and    acetic acetate followed by treatment with sodium hydroxide;    alternatively can be used sodium nitrite in a mixture of acetic    anhydride acetic acid, followed by treatment with sodium hydroxide;    alternatively the acetamide group of compound of Formula 10 can be    converted into the primary amine group by reacting with hydrazine or    with Boc₂O, DMAP followed by hydrazine; such primary amine can be    converted into the corresponding hydroxyl group (compound of    Formula 11) by an oxidative conversion of the primary amine into the    corresponding aldehyde with 4-formyl-1-methylpyridinium    benzenesulphonate or other pyridinium ion, followed by DBU or other    base treatment and further hydrolization, and followed by the    reduction of the aldehyde to the corresponding hydroxyl group with    lithium aluminium hydride or other reducing agent;-   forming the protected compound of Formula 26 by reacting with    t-butyldiphenylsilyl chloride and dimethylaminopyridine in    dichloromethane;-   transforming the silylated compound of Formula 26 into the    intermediate II by deprotection of the OCH₂R protecting group, by    reacting under reductive conditions or acid conditions.    Typical procedures are with palladium black under hydrogen    atmosphere, or aqueous TFA, or tributyltin hydride and dichlorobis    (triphenylphosphine palladium).

In yet another preferred modification, the cyano compound of Formula 2can be transformed into Intermediate 11 using an extension of the schemeII, involving the further steps of.

-   formation of the protected hydroxy compound of Formula 26 by    reacting 25 with tert-butyldiphenylsilyl chloride in the presence of    a base;-   final cleavage of the allyl group with tributyltin hydride and    dichloropalladium-bis (triphenylphosphine) in 26 that leads to the    formation of the intermediate 11.

Thus, it is possible to transform cyanosafracin B into a number ofintermediates and derivatives with potential antitumor therapeuticactivity. These intermediates can be made starting from alreadydescribed compounds, or using alternative routes.

Intermediates described herein comprise compound 47, and a numbers ofamide derivatives made using compounds 45 or 43.

In Scheme VIII is described formation of compound 47 using the followingsequence:

-   forming the thiourea compound of Formula 3 by reacting compound of    Formula 2 with phenyl isothiocyanate;-   converting the thiourea compound of Formula 3 into the acetamide of    Formula 5 by an hydrolysis in acid media followed by addition of    acetic anhydride; The intermediate amine compound of Formula 4 can    be isolated by quenching the hydrolysis in acid media with sodium    bicarbonate, but this intermediate is highly unstable, and is    transformed quickly into a five member cyclic imine, named compound    6;-   forming the protected compound of Formula 7 by reacting with    bromomethylmethyl ether and diisopropylethylamine in    dichloromethane;-   selectively de-methylating the methoxy group of the quinone system    of compound of Formula 7 into the compound of Formula 8 by reacting    with methanolic solution of sodium hydroxide;-   transforming the compound of Formula 8 into methylenedioxy-compound    of Formula 10 by the preferred following sequence: (1) quinone group    of compound 8 is reduced with 10% Pd/C under hydrogen    atmosphere; (2) the hydroquinone intermediate is converted into the    methylene-dioxy compound of Formula 9 by reacting with    bromochloromethane and cesium carbonate under hydrogen    atmosphere; (3) compound of Formula 9 is transformed into compound    of Formula 10 by protecting the free hydroxyl group as a allyloxy    group, by reacting with allyl-bromide and cesium carbonate;-   transforming the compound of formula 9 into acetyl-derivative 46 by    reaction with acetyl chloride in pyridine;-   transforming compound of formula 46 into de-protected compound 47 by    reaction with hydrochloric acid in dioxane.

Other useful amide intermediate derivatives are made starting fromalready described intermediate 45 using the next scheme:

The second step is optional. This process is an important part of theinvention, particularly where the group R is a group R^(a) as previouslydefined. Furthermore, the Scheme VIII can be readily broadened to enablepreparation of compounds of formula (XXIII), by inclusion in thestarting material of a different group at the 5-position, either a groupdirectly intended for the product or a group which can be removed orotherwise modified to give the desired group.

From compound 45 can be made a group of analogs through the followingsequence:

-   acylation in the amino group of compound of Formula 45 by a wide    range of acyl derivatives to provide the corresponding amides, where    preferred acyl groups are acetyl, cinnamoyl chloride,    p-trifluorocinnamoyl chloride, isovaleryl chloride    phenylisothiocyanate or aminoacids, or the other examples previously    given of groups R^(a)CO—.-   transforming the CN group into an OH group by reaction with silver    nitrate in a mixture AcN/H₂O.

Other useful amide intermediate derivatives are made starting fromalready described intermediate 43 using the next scheme:

From Compound 43 can be obtained another group of interestingderivatives using the following sequence:

-   (a) acylation in the amino group of compound of Formula 43 by a wide    range of acyl derivatives to provide the corresponding amides, where    preferred acyl groups are acetyl, cinnamoyl chloride,    p-trifluorocinnamoyl chloride, isovaleryl chloride or aminoacids, or    the other examples previously given of groups R^(a)CO—.-   (b) transforming the CN group into an OH group by reaction with    silver nitrate in a mixture AcN/H₂O

Reflecting the active compounds, an important process in accordance withthis invention is as follows:

-   where R⁵ for the end product is as defined for the compound (XXII)    and may be different in the starting material and converted thereto    as part of the process,-   R¹⁸ is a hydroxy group in the end product but may be a protected    hydroxy group in the starting material and converted thereto as part    of the process,-   R¹² for the end product may be the same as in the starting material    or may be converted thereto as part of the process,-   R²¹ for the end product is as defined and if a hydroxy group may be    formed from a cyano group as part of the process,-   R^(a) is as defined, and may be further acylated as part of the    process to give an end product with an acylated R^(a) group as    discussed.

R⁵ is preferably oxyacetyl or other small oxyacyl group in the startingmaterial and is not changed in the reaction. R¹⁸ is preferably a hydroxygroup in the starting material and is not changed in the reaction. R¹²is preferably —NCH₃— in the starting material and is not changed in thereaction. R²¹ the end product is as defined and if a hydroxy group maybe formed from a cyano group as part of the process. R^(a) is in thefinal product is preferably as defined in relation to the compound offormula (XXIII).

Another important method of this invention includes the reaction:

Another important method of this invention includes the reaction:

Another important method of this invention includes the reactionincludes the reaction where a group R¹ is aminomethylene is converted toa hydroxyethylene group.

Another important method of this invention includes the reaction forpreparing a 21-cyano compound of formula (XVI) which comprises reactinga compound of formula (XV):

where R¹, R⁵, R⁸, R^(14a), R^(14b), R¹⁵ and R¹⁸ are as defined and R²¹is a hydroxy group, with a source of cyanide ion, to give the desired21-cyano compound.

In addition, processes using other nucleophile-containing compounds, toproduce similar compounds of formula (XVI) wherein the 21-position isprotected by another nucleophilic group, a 21-Nuc group, are alsoenvisaged. For example, a 21-Nuc compound of formula (XVI) with analkylamino substituent at the 21-position can be produced by reactingthe compound of formula (XV) wherein R²¹ is a hydroxy group with asuitable alkylamine. A 21-Nuc compound of formula (XVI) with analkylthio substituent at the 21-position can also be produced byreacting the compound of formula (XV) wherein R²¹ is a hydroxy groupwith a suitable alkanethiol. Alternatively, a 21-Nuc compound of formula(XVI) with an α-carbonylalkyl substituent at the 21-position can beproduced by reacting the compound of formula (XV) wherein R²¹ is ahydroxy group with a suitable carbonyl compound, typically in thepresence of a base. Other synthetic routes are available for other21-Nuc compounds.

Another important reaction of this invention involves treatment of a21-cyano product of this invention to form a 21-hydroxy compound. Suchcompounds have interesting in vivo properties.

For the avoidance of doubt, the stereochemistries indicated in thispatent specification are based on our understanding of the correctstereochemistry of the natural products. To the extent that an error isdiscovered in the assigned stereochemistry, then the appropriatecorrection needs to be made in the formulae given throughout in thispatent specification. Furthermore, to the extent that the syntheses arecapable of modification, this invention extends to stereoisomers.

Cytotoxic Activitiy

IC₅₀(μM) Compound P-388 A-549 HT-29 MEL-28 CV-1 DU-145

2 0.009 0.018 0.018 0.018 0.023

14 0.15 >0.15 0.15 >0.15

15 1.44 1.44 1.44 1.44

16 >1.5 >1.5 >1.5 >1.5

17 1.4 1.4 1.4 1.4

18 0.01 0.01 0.01 0.01

19 0.08 0.16 0.01 0.16

20 0.01 0.01 0.01 0.01

21 0.019 0.019 0.019 0.019

22 0.014 0.014 0.014 0.014 0.014 0.014

23 0.13 0.13 0.13 0.13 0.13 0.13

24 0.18 1.8 1.8 1.8 1.8 1.8

25 0.2 0.2 0.2 0.2 0.2

35 0.008 0.008 0.008 0.008

36 0.01 0.01 0.01 0.01

28 0.001 0.001 0.001 0.001 0.001 0.001

42 0.13 0.13 0.13 0.13 0.13

43 0.008 0.016 0.008 0.008 0.016

44 0.001 0.001 0.001 0.001 0.001

45 0.01 0.01 0.01 0.01 0.01

3 0.015 0.015 0.015 0.015 0.018

6 2.171 2.171 2.171 2.171 2.171

5 0.005 0.005 0.005 0.005

7 0.22 0.22 0.22 0.22 0.22

8 >9 >18.1 >18.1 >18.1 >18.1

9 >1.77 >1.77 >1.77 >1.77 >1.77

10 >1.65 >1.65 >1.65 >1.65 >1.65

46 0.016 0.016 0.016 0.016 0.016

47 0.001 0.001 0.001 0.001 0.001

48 0.0008 0.001 0.0008 0.0008 0.001

49 0.007 0.007 0.007 0.007 0.007

50 0.0001 0.0001 0.0001 0.0001 0.0001

51 0.0001 0.0001 0.0001 0.0001 0.0001

52 0.001 0.001 0.001 0.001 0.001

53 0.0001 0.0001 0.0001 0.0001 0.0001

54 0.001 0.001 0.001 0.001 0.001

55 0.01 0.01 0.01 0.01 0.01

56 0.18 0.9 0.18 0.8 0.9

57 0.14 0.14 0.14 0.14 0.14

58 0.001 0.001 0.001 0.001 0.001

60 0.001 0.001 0.0001 0.001 0.0005

61 0.001 0.001 0.001 0.001 0.001

62 0.001 0.001 0.0005 0.001

63 0.0001 0.0001 0.0001 0.0001 0.0001

64 0.001 0.001 0.001 0.001

65 0.0001 0.0001 0.0001 0.0001 0.0001

66 0.0001 0.0001 0.0001 0.0001 0.0001

67 0.0001 0.0001 0.0001 0.0001 0.0001

68 0.0008 0.001 0.0008 0.0008 0.001

69 0.001 0.001 0.001 0.001 0.001

70 0.0001 0.0001 0.0001 0.0001 0.0001

71 0.0008 0.0008 0.0001 0.0008 0.0001

72 0.0001 0.0001 0.0001 0.0001 0.0001

73 0.0001 0.0001 0.0001 0.0001 0.0001

74 0.0001 0.0001 0.0001 0.0001 0.0001

75 0.1 0.1 0.1 0.1 0.1

76 0.1 0.1 0.1 0.1 0.1

77 0.0001 0.0001 0.0001 0.0001 0.0001 0.0001

78 0.0001 0.0008 0.0001 0.0001 0.0008

79 0.001 0.001 0.001 0.001 0.001

80 0.0001 0.0001 0.0001 0.0001 0.0001

81 0.0007 0.0007 0.0007 0.0007 0.0007

82 0.0001 0.0001 0.0001 0.0001 0.0001

83 0.0001 0.0001 0.0001 0.0001 0.0001 0.0001

84 0.0001 0.0008 0.0001 0.0001 0.0008

85 0.0006 0.001 0.0006 0.001 0.0006

86 0.001 0.001 0.001 0.001 0.001 0.001

87 0.0001 0.0001 0.0001 0.0001 0.0001

88 0.0007 0.0007 0.0007 0.0007 0.0007

89 0.001 0.007 0.001 0.001 0.007

90 0.01 0.01 0.01 0.01 0.01

91 0.001 0.001 0.001 0.001 0.001

92 0.0001 0.0001 0.0001 0.0001 0.0001

93 0.001 0.001 0.001 0.001 0.001 0.001

94 0.0007 0.0007 0.0007 0.0007 0.0007

95 0.0001 0.0001 0.0001 0.0001 0.0001

96 0.001 0.007 0.001 0.001 0.007

97 >1 >1 >1 >1 >1

98 >1 >1 >1 >1 >1

99 0.7 0.7 0.7 0.7 0.7 0.7

100 0.1 0.1 0.1 0.1 0.1

101 0.1 0.1 0.1 0.1 0.1

102 0.1 0.1 0.1 0.1 0.1

103 0.1 0.1 0.1 0.1 0.1

104 0.1 0.1 0.1 0.1 0.1

105 0.1 0.1 0.1 0.1 0.1

106 0.6 0.6 0.6 0.6 0.6

107 0.1 0.1 0.1 0.1 0.1

108 0.01 0.07 0.07 0.07 0.07

109 0.0001 0.0008 0.0008 0.0008 0.0008

110 0.001 0.001 0.001 0.001 0.001

111 0.0001 0.0001 0.0001 0.0001 0.0001

112 0.0007 0.0007 0.0007 0.0007 0.0007

113 0.0001 0.0001 0.0001 0.0001 0.0001

114 0.0001 0.0001 0.0001 0.0001 0.0001

115 0.0001 0.0001 0.0001 0.0001 0.0001

116 0.0001 0.0007 0.0007 0.0007 0.0007

117 0.06 0.06 0.06 0.06 0.06

118 0.001 0.001 0.001 0.001 0.001

119 0.001 0.001 0.001 0.001 0.001

120 0.06 0.06 0.06 0.06 0.06

121 0.006 0.006 0.006 0.006 0.006

122 0.1 0.1 0.1 0.1 0.1

124 0.0001 0.0001 0.0001 0.0001 0.0001

125 0.001 0.001 0.001 0.001 0.001

126 0.0001 0.0001 0.0001 0.0001 0.0001

127 0.0001 0.0001 0.0001 0.0001 0.0001

128 0.0001 0.0001 0.0001 0.0001 0.0001

129 0.1 0.1 0.1 0.1 0.1

130 0.1 0.1 0.1 0.1 0.1

131 0.5 0.5 0.5 0.5 0.5

132 0.1 0.1 0.1 0.1 0.1

133 0.05 0.05 0.05 0.05 0.05

134 0.5 0.5 0.5 0.5 0.5 0.5

135 0.01 0.01 0.01 0.01 0.01

136 0.001 0.001 0.001 0.001 0.001

137 0.01 0.01 0.01 0.01 0.01

138 0.006 0.006 0.006 0.006 0.006

139 0.01 0.01 0.01 0.01 0.01

140 0.08 0.08 0.08 0.08 0.08

141 0.01 0.01 0.01 0.01 0.01 0.01

174 0.0013 0.0013

175 0.007 0.007

176 0.014 0.014

177 >1 >1

178 0.00012 0.00012

179 0.012 0.012

180 0.00015 0.00015

181 0.00015 0.00015

182 0.0015 0.0015

183 0.013 0.013

184 0.0015 0.0015

185 0.12 0.12

186 0.0014 0.0014

187 0.013 0.013

188 0.012 0.012

189 0.06 0.06

190 0.013 0.013

191 0.13 0.13

192 0.12 0.12

193 0.11 0.11

194 0.012 0.012

195 0.012 0.012

196 0.1 0.1

197 0.0018 0.0018

198 0.0015 0.0015

199 >1 >1

202 0.056 0.056Cytotoxic Activity (M)

               SOLIDTUMORS                 LINE

Bladder 5637 6.02E−10 3.42E−10 1.91E−10 2.04E−11 Breast MX-1 1.65E−06 NA2.38E−09 NA Colon HT-29 7.84E−10 1.97E−08 2.12E−09 8.44E−12 GastricHs746t 7.90E−12 2.18E−09 7.10E−11 2.21E−09 Liver SK-HEP-1 1.79E−126.01E−11 3.15E−09 9.91E−11 NSCL A549 3.25E−09 7.68E−06 NA NA OvarySK-OV-3 4.39E−11 1.02E−07 8.74E−09 NA Pancreas PANC-1 7.22E−11 4.17E−091.29E−10 1.19E−10 Pharnynx FADU 5.41E−11 1.58E−09 3.71E−10 5.98E−09Prostate PC3 6.65E−09 2.15E−09 4.70E−09 1.52E−10 Prostate DU-1455.73E−10 1.83E−07 2.22E−09 NA Prostate LNCAP 5.45E−10 2.17E−10 3.94E−11Renal 786-O 6.58E−12 1.59E−09 1.72E−09 1.03E−10 SCL NCI-H187 7.14E−149.57E−10 7.78E−14 Retinoblastoma Y-79 7.14E−14 7.36E−10 8.85E−11Melanoma Mel-28 2.60E−10 3.17E−09 2.18E−09 1.23E−10 Fibrosarcoma SW-6949.91E−10 NA 1.39E−06 NA Chondrosarcoma CHSA 3.24E−10 6.77E−09 1.39E−092.30E−10 Osteosarcoma OSA-FH 1.94E−09 1.39E−09 1.09E−09 1.11E−10                   SOLIDTUMORS                     LINE

  

Bladder 5637 1.65E−10 7.85E−10 3.18−09 Breast MX-1 NA 2.85E−06 NA ColonHT-29 7.43E−10 1.2E−10 NA Gastric Hs746t 9.35E−10 6.25E−09 1.37E−07Liver SK-HEP-1 1.40E−09 9.03E−10 9.50E−09 NSCL A549 NA NA NA OvarySK-OV-3 NA NA Pancreas PANC-1 8.93E−10 2.58E−9 1.03E−08 Pharnynx FADU8.41E−10 3.77E−08 1.14E−09 Prostate PC3 8.13E−10 9.34E−09 ProstateDU-145 NA NA NA Prostate LNCAP NA Renal 786-O 7.88E−10 2.90E−09 1.00E−08SCL NCI-H187 2.07E−12 Retinoblastoma Y-79 1.31E−11 7.78E−09 MelanomaMel-28 1.08E−09 1.13E−09 4.48E−09 Fibrosarcoma SW-694 NA ChondrosarcomaCHSA 1.08E−09 2.25E−09 1.09E−08 Osteosarcoma OSA-FH 8.84E−10 1.35−089.50E−09               LEUKEMIAS&LYMPHOMAS                 LINE  

ALL HL60 9.38E−09 Promyelocytic leukemia ALL Molt 3 6.13E−10 2.8E−095.66E−10 1.55E−14 Acute lymphoblastic CML K562 2.33E−07 Chronicmyelogenous Leukemia Mo-B Hairy B-cell Lymphoma T- H9 1.99E−11 cellLymphoma Hut 78 5.50E−11 2.57E−10 4.62E−09 6.21E−11 Cutaneus T cellLymphoma MC116 2.15E−10 2.65E−10 3.8E−09 NA undifferentiated LymphomaRAMOS 7.77E−13 Burkitts B cell Lymphoma U-937 1.77E−10 5.27E−11 3.28E−113.06E−11 Histiocytic                   LEUKEMIAS&LYMPHOMAS                    LINE

  

ALL HL60 5.92E−09 1.23E−10 3.97E−10 Promyelocytic leukemia ALL Molt 37.53E−12 8.85E−10 2.54E−09 Acute lymphoblastic CML K562 1.09E−084.45E−08 Chronic myelogenous Leukemia Mo-B Hairy B-cell Lymphoma T- H94.48E−09 1.14E−08 cell Lymphoma Hut 78 9.9E−10 1.06E−08 7.46E−09Cutaneous T Cell Lymphoma MC116 NA 1.41E−09 1.13E−08 undifferentiatedLymphoma RAMOS 5.26−11 8.85E−10 7.15E−09 Burkitts B cell lymphoma U-9375.15E−10 Histiocytic                  SOLIDTUMORS                   LINE

Bladder 5637 2.81E−09 2.84E−10 Breast MX-1 2.50E−06 NA Colon HT-29 NA8.97E−09 Gastric Hs746t 2.97E−08 9.19E−09 Liver SK-HEP-1 5.07E−091.08E−09 NSCL A549 NA 9.41E−09 Ovary SK-OV-3 2.21E−07 NA Pancreas PANC-12.90E−09 1.00E−09 Pharnynx FADU 7.94E−09 1.39E−08 Prostate PC3 1.46−089.32E−10 Prostate DU-145 NA NA Prostate LNCAP 5.39E−09 Renal 786-O6.55E−09 1.72E−09 SCL NCI-H187 3.98E−11 Retinoblastoma Y-79 3.14E−09Melanoma Mel-28 3.05E−08 1.15E−09 Fibrosarcoma SW-694 NA NAChondrosarcoma CHSA 1.73E−08 2.10E−09 Osteosarcoma OSA-FH 8.56E−081.30E−09                   SOLIDTUMORS                    LINE   

Bladder 5637 9.91E−10 1.17E−09 Breast MX-1 NA 1.92E−09 Colon HT-29 NA NAGastric Hs746t 1.36E−09 8.15E−09 Liver SK-HEP-1 1.17E−09 6.21E−09 NSCLA549 NA NA Ovary SK-OV-3 2.90E−08 NA Pancreas PANC-1 1.37E−09 8.61E−09Pharnynx FADU 3.05E−08 4.38E−08 Prostate PC3 Prostate DU-145 NA NAProstate LNCAP 2.38E−08 1.77E−08 Renal 786-O 2.27E−09 1.54E−08 SCLNCI-H187 2.41E−11 9.89E−11 Retinoblastoma Y-79 3.08E−10 7.45E−10Melanoma Mel-28 2.85E−09 1.42E−08 Fibrosarcoma SW-694 ChondrosarcomaCHSA 1.63E−09 2.91E−08 Osteosarcoma OSA-FH 4.37E−09 1.15E−08                LEUKEMIAS&LYMPHOMAS                   LINE

ALL HL60 1.50E−08 Promyelocytic leukemia ALL Molt 3 1.62E−09 3.87E−09Acute lymphoblastic CML K562 6.89E−08 Chronic myelogenous LymphomaT-cell H9 1.08E−08 Lymphoma Hut 78 7.33E−09 1.97E−09 Cutaneous T cellLymphoma MC116 1.62E−08 3.81E−09 undifferentiated Lymphoma RAMOS 1.1E−09Burkitts B cell Lymphoma U-937 1.92E−09 1.08E−09 Histiocytic                 LEUKEMIAS&LYMPHOMAS                    LINE   

ALL HL60 4.93E−10 7.36E−09 Promyelocytic leukemia ALL Molt 3 9.86E−109.86E−10 Acute lymphoblastic CML K562 1.87E−08 1.18E−08 Chronicmyelogenous Lymphoma T-cell H9 1.20E−08 2.43−08 Lymphoma Hut 78 CutaneusT cell Lymphoma MC116 1.04E−09 1.49E−09 undifferentiated Lymphoma RAMOS5.01E−09 Burkitts B cell Lymphoma U-937 Histiocytic                  SOLIDTUMORS                    LINE

Bladder 5637 1.14E−08 1.71E−08 Breast MX-1 2.81E−08 7.25E−13 Colon HT-294.08E−07 2.96E−07 Gastric Hs746t 3.57E−08 1.24E−09 Liver SK-HEP-11.63−08 1.94E−09 NSCL A549 2.81E−06 1.56−05 Ovary SK-OV-3 7.03E−067.78E−08 Pancreas PANC-1 1.03E−08 9.47E−09 Pharnynx FADU 4.59E−072.46E−08 Prostate PC3 7.88E−08 Prostate DU-145 7.03E−08 1.56E−06Prostate LNCAP 5.98E−07 6.83E−08 Renal 786-O 1.46E−08 5.26E−12 SCLNCI-H187 8.02E−10 7.78E−14 Retinoblastoma Y-79 8.85E−10 7.78E−14Melanoma Mel-28 1.76E−08 5.89E−08 Fibrosarcoma SW-694 3.38E−06 6.69E−06Chondrosarcoma CHSA 2.53E−08 4.49E−08 Osteosarcoma OSA-FH 6.34E−085.26E−07                    SOLIDTUMORS                     LINE

  

Bladder 5637 7.88E−10 3.02E−08 Breast MX-1 NA 4.75E−08 Colon HT-298.99E−09 1.34E−08 Gastric Hs746t 2.95E−08 7.05E−07 Liver SK-HEP-11.29E−09 6.12E−08 NSCL A549 8.22E−06 8.49E−09 Ovary SK-OV-3 3.55E−08Pancreas PANC-1 5.68E−10 1.28E−08 Pharnynx FADU 5.40E−11 2.47E−08Prostate PC3 7.71E−10 6.18E−10 Prostate DU-145 NA 1.17E−08 ProstateLNCAP 3.29E−07 Renal 786-O 9.23E−10 1.13E−08 SCL NCI-H187 2.33E−10Retinoblatoma Y-79 1.03E−08 2.64E−09 Melanoma Mel-28 2.23E−08 1.25E−08Fibrosarcoma SW-694 8.53E−06 NA Chondrosarcoma CHSA 1.55E−05 2.95E−08Osteosarcoma OSA-FH 1.29E−09 5.01E−08                  LEUKEMIAS&LYMPHOMAS                     LINE

ALL HL60 1.34E−08 Promyelocytic leukemia ALL Molt 3 1.44E−08 2.48E−09Acute lymphoblastic CML K562 1.56E−07 6.13E−08 Chronic myelogenousLymphoma T-cell H9 1.56E−07 1.91E−08 Lymphoma Hut 78 6.47E−08 7.31E−09Cutaneous T cell Lymphoma MC116 1.69E−08 6.38E−09 undifferentiatedLymphoma RAMOS 8.86E−09 7.15E−10 Burkitts B cell Lymphoma U-937 7.6E−08Histiocytic                  LEUKEMIAS&LYMPHOMAS                    LINE

 

ALL HL60 3.1E−09 Promyelocytic leukemia ALL Molt 3 8.69E−11 4.63E−08Acute lymphoblastic CML K562 2.11E−08 Chronic myelogenous LymphomaT-cell H9 2.17E−08 6.76E−08 Lymphoma Hut 78 4.81E−08 2.06E−08 CutaneousT cell Lymphoma MC116 5.27E−11 1.51E−08 undifferentiated Lymphoma RAMOS1.86E−09 9.09E−09 Burkitts B cell Lymphoma U-937 1.03E−08 Histicytic

EXAMPLES OF THE INVENTION

The present invention is illustrated by the following examples.

Example 1

To a solution of 2 (21.53 g, 39.17 ml) in ethanol (200 ml),tert-butoxycarbonyl anhydride (7.7 g, 35.25 ml) was added and themixture was stirred for 7 h at 23° C. Then, the reaction wasconcentrated in vacuo and the residue was purified by flash columnchromatography (SiO₂, hexane:ethyl acetate 6:4) to give 14 (20.6 g 81%)as a yellow solid.

Rf: 0.52 (ethyl acetate:CHCl₃ 5:2).

¹H NMR (300 MHz, CDCl₃): δ 6.49 (s, 1H), 6.32 (bs, 1H) 5.26 (bs, 1H),4.60 (bs, 1H), 4.14 (d, J=2.4 Hz, 1H), 4.05 (d, J=2.4 Hz, 1H), 3.94 (s,3H), 3.81 (d, J=4.8 Hz, 1H), 3.7 (s, 3H), 3.34 (br d, J=7.2 Hz, 1H),3.18–3.00 (m, 5H), 2.44 (d, J=18.3 Hz, 1H), 2.29 (s, 3H), 2.24 (s, 3H),1.82 (s, 3H), 1.80–1.65 (m, 1H), 1.48 (s, 9H), 0.86 (d, J=5.7 Hz, 3H)

¹³C NMR (75 MHz, CDCl₃): δ 185.5, 180.8, 172.7, 155.9, 154.5, 147.3,143.3, 141.5, 135.3, 130.4, 129.2, 127.5, 120.2, 117.4, 116.9, 80.2,60.7, 60.3, 58.5, 55.9, 55.8, 54.9, 54.4, 50.0, 41.6, 40.3, 28.0, 25.3,24.0, 18.1, 15.6, 8.5.

ESI-MS m/z: Calcd. for C₃₄H₄₃N₅O₈: 649.7. Found (M+H)⁺: 650.3.

Example 2

To a stirred solution of 14 (20.6 g, 31.75 ml) in CH₃CN (159 ml),diisopropylethylamine (82.96 ml, 476.2 ml), methoxymethylene bromide(25.9 ml, 317.5 ml) and dimethylaminopyridine (155 mg, 1.27 ml) wereadded at 0° C. The mixture was stirred at 23° C. for 24 h. The reactionwas quenched at 0° C. with aqueous 0.1N HCl (750 ml) (pH=5), andextracted with CH₂Cl₂ (2×400 ml). The organic phase was dried (sodiumsulphate) and concentrated in vacuo. The residue was purified by flashcolumn chromatography (SiO₂, gradient hexane:ethyl acetate 4:1 tohexane:ethyl acetate 3:2) to give 15 (17.6 g, 83%) as a yellow solid.

Rf: 0.38 (hexane:ethyl acetate 3:7).

¹H NMR (300 MHz, CDCl₃): δ 6.73 (s, 1H), 5.35 (bs, 1H), 5.13 (s, 2H),4.50 (bs, 1H), 4.25 (d, J=2.7 Hz, 1H), 4.03 (d, J=2.7 Hz, 1H), 3.97 (s,3H), 3.84 (bs, 1H), 3.82–3.65 (m, 1H), 3.69 (s, 3H), 3.56 (s, 3H),3.39–3.37 (m, 1H), 3.20–3.00 (m, 5H), 2.46 (d, J=18 Hz, 1H), 2.33 (s,3H), 2.23 (s, 3H), 1.85 (s, 3H), 1.73–1.63 (m, 1H), 1.29 (s, 9H), 0.93(d, J=5.1 Hz, 3H)

¹³C NMR (75 MHz, CDCl₃): δ 185.4, 180.9, 172.4, 155.9, 154.5, 149.0,148.4, 141.6, 135.1, 131.0, 129.9, 127.6, 124.4, 123.7, 117.3, 99.1,79.3, 60.7, 59.7, 58.4, 57.5, 56.2, 55.9, 55.0, 54.2, 50.0, 41.5, 39.9,28.0, 25.2, 24.0, 18.1, 15.6, 8.5.

ESI-MS m/z: Calcd. for C₃₆H₄₇N₅O₉: 693.8. Found (M+H)⁺: 694.3.

Example 3

To a flask containing 15 (8 g, 1.5 ml) in methanol (1.61) an aqueoussolution of 1M sodium hydroxide (3.2 l) was added at 0° C. The reactionwas stirred for 2 h at this temperature and then, quenched with 6M HClto pH=5. The mixture was extracted with ethyl acetate (3×1 l) and thecombined organic layers were dried over sodium sulphate and concentratedin vacuo. The residue was purified by flash column chromatography (SiO₂,gradient CHCl₃ to CHCl₃:ethyl acetate 2:1) to afford 16 (5.3 mg, 68%).

Rf: 0.48 (CH₃CN:H₂O 7:3, RP-C18)

¹H NMR (300 MHz, CDCl₃): δ 6.73 (s, 1H), 5.43 (bs, 1H), 5.16 (s, 2H),4.54 (bs, 1H), 4.26 (d, J=1.8 Hz, 1H), 4.04 (d, J=2.7 Hz 1H), 3.84 (bs,1H), 3.80–3.64 (m, 1H), 3.58 (s, 3H), 3.41–3.39 (m, 1H), 3.22–3.06 (m,5H), 2.49 (d, J=18.6 Hz 1H) 2.35 (s, 3H), 2.30–2.25 (m, 1H), 2.24 (s,3H), 1.87 (s, 3H), 1.45–1.33 (m, 1H), 1.19 (s, 9H), 1.00 (br d, J=6.6 Hz3H)

¹³C NMR (75 MHz, CDCl₃): δ 184.9, 180.9, 172.6, 154.7, 151.3, 149.1,148.6, 144.7, 132.9, 131.3, 129.8, 124.5, 123.7, 117.3, 116.8, 99.1,79.4, 59.8, 58.6, 57.7, 56.2, 55.6, 54.9, 54.5, 50.1, 41.6, 40.1, 28.0,25.3, 24.4, 18.1, 15.7, 8.0.

ESI-MS m/z: Calcd. for C₃₅H₄₅N₅O₉: 679.7. Found (M+H)⁺: 680.3

Example 4

To a degassed solution of compound 16 (1.8 g, 2.64 ml) in DMF (221 ml)10% Pd/C (360 mg) was added and stirred under H₂ (atmospheric pressure)for 45 min. The reaction was filtered through celite under argon, to aflask containing anhydrous Cs₂CO₃ (2.58 g, 7.92 ml). Then,bromochloromethane (3.40 ml 52.8 ml), was added and the tube was sealedand stirred at 100° C. for 2 h. The reaction was cooled, filteredthrough a pad of celite and washed with CH₂Cl₂. The organic layer wasconcentrated and dried (sodium sulphate) to afford 17 as a brown oilthat was used in the next step with no further purification.

Rf: 0.36 (hexane:ethyl acetate 1:5, SiO₂).

¹H NMR (300 MHz, CDCl₃): δ 6.68 (s, 1H), 6.05 (bs, 1H), 5.90 (s, 1H),5.79 (s, 1H), 5.40 (bs, 1H), 5.31–5.24 (m, 2H), 4.67 (d, J=8.1 Hz, 1H),4.19 (d, J=2.7 Hz, 1H), 4.07 (bs, 1H), 4.01 (bs, 1H), 3.70 (s, 3H), 3.67(s, 3H), 3.64–2.96 (m, 5H), 2.65 (d, J=18.3 Hz, 1H), 2.33 (s, 3H), 2.21(s, 3H), 2.04 (s, 3H), 2.01–1.95 (m, 1H), 1.28 (s, 9H), 0.87 (d, J=6.3Hz, 3H)

¹³C NMR (75 MHz, CDCl₃): δ 172.1, 162.6, 154.9, 149.1, 145.7, 135.9,130.8, 130.7, 125.1, 123.1, 117.8, 100.8, 99.8, 76.6, 59.8, 59.2, 57.7,57.0, 56.7, 55.8, 55.2, 49.5, 41.6, 40.1, 36.5, 31.9, 31.6, 29.7, 28.2,26.3, 25.0, 22.6, 18.2, 15.8, 14.1, 8.8.

ESI-MS m/z: Calcd. for C₃₆H₄₇N₅O₉: 693.34. Found (M+H)⁺: 694.3.

Example 5

To a flask containing a solution of 17 (1.83 g, 2.65 ml) in DMF (13 ml).Cs₂CO₃ (2.6 g, 7.97 ml), and allyl bromide (1.15 ml, 13.28 ml) wereadded at 0° C. The resulting mixture was stirred at 23° C. for 1 h. Thereaction was filtered through a pad of celite and washed with CH₂Cl₂.The organic layer was dried and concentrated (sodium sulphate). Theresidue was purified by flash column chromatography (SiO₂, CHCl₃:ethylacetate 1:4) to afford 18 (1.08 mg, 56%) as a white solid.

Rf: 0.36 (CHCl₃:ethyl acetate 1:3).

¹H NMR (300 MHz, CDCl₃): δ 6.70 (s, 1H), 6.27–6.02 (m, 1H), 5.94 (s,1H), 5.83 (s, 1H), 5.37 (dd, J₁=1.01 Hz, J₂=16.8 Hz, 1H), 5.40 (bs, 1H),5.25 (dd, J₁=1.0 Hz, J₂=10.5 Hz, 11H), 5.10 (s, 2H), 4.91 (bs, 1H),4.25–4.22 (m, 1H), 4.21 (d, J=2.4 Hz, 1H), 4.14–4.10 (m, 1H), 4.08 (d,J=2.4 Hz, 1H), 4.00 (bs, 1H), 3.70 (s, 3H), 3.59 (s, 3H), 3.56–3.35 (m,2H), 3.26–3.20 (m, 2H), 3.05–2.96 (dd, J₁=8.1 Hz, J₂=18 Hz, 1H), 2.63(d, J=18 Hz, 1H), 2.30 (s, 3H), 2.21 (s, 3H), 2.09 (s, 3H), 1.91–1.80(m, 1H), 1.24 (s, 9H), 0.94 (d, J=6.6 Hz, 3H)

¹³C NMR (75 MHz, CDCl₃): δ 172.0, 154.8, 148.8, 148.6, 148.4, 144.4,138.8, 133.7, 130.9, 130.3, 125.1, 124.0, 120.9, 117.8, 117.4, 112.8,112.6, 101.1, 99.2, 73.9, 59.7, 59.3, 57.7, 56.9, 56.8, 56.2, 55.2,40.1, 34.6, 31.5, 28.1, 26.4, 25.1, 22.6, 18.5, 15.7, 14.0, 9.2.

ESI-MS m/z: Calcd. for C₃₉H₅₁N₅O₉: 733.4. Found (M+H)⁺: 734.4.

Example 6

To a solution of 18 (0.1 g, 0.137 ml) in dioxane (2 ml), 4.2MHCl/dioxane (1.46 ml) was added and the mixture was stirred for 1.2 h at23° C. The reaction was quenched at 0° C. with sat. Aqueous sodiumbicarbonate (60 ml) and extracted with ethyl acetate (2×70 ml). Theorganic layers were dried (sodium sulphate) and concentrated in vacuo toafford 19 (267 mg, 95%) as a white solid that was used in subsequentreactions with no further purification.

Rf: 0.17 (ethyl acetate:methanol 10:1, SiO₂)

¹H NMR (300 MHz, CDCl₃): δ 6.49 (s, 1H), 6.12–6.00 (m, 1H), 5.94 (s,1H), 5.86 (s, 1H), 5.34 (dd, J=1.0 Hz, J=17.4 Hz, 1H), 5.25 (dd, J=1.0Hz, J=10.2 Hz, 1H), 4.18–3.76 (m, 5H), 3.74 (s, 3H), 3.71–3.59 (m, 1H),3.36–3.20 (m, 4H), 3.01–2.90 (m, 1H), 2.60 (d, J=18.0 Hz, 1H), 2.29 (s,3H), 2.24 (s, 3H), 2.11 (s, 3H), 1.97–1.86 (m, 1H), 0.93 (d, J=8.7 Hz,3H)

¹³C NMR (75 MHz, CDCl₃): δ 175.5, 148.4, 146.7, 144.4, 142.4, 138.9,133.7, 131.3, 128.3, 120.8, 117.9, 117.4, 113.8, 112.4, 101.1, 74.2,60.5, 59.1, 56.5, 56.1, 56.3, 56.0, 55.0, 50.5, 41.6, 39.5, 29.5, 26.4,24.9, 21.1, 15.5, 9.33.

ESI-MS m/z: Calcd. for C₃₂H₃₉N₅O₆: 589. Found (M+H)⁺: 590.

Example 7

To a solution of 19 (250 mg, 0.42 ml) in CH₂Cl₂ (1.5 ml), phenylisothiocyanate (0.3 ml, 2.51 ml) was added and the mixture was stirredat 23° C. for 1 h. The reaction was concentrated in vacuo and theresidue was purified by flash column chromatography (SiO₂, gradientHexane to 5:1 hexane:ethyl acetate) to afford 20 (270 mg, 87%) as awhite solid.

Rf: 0.56 (CHCl₃:ethyl acetate 1:4).

¹H NMR (300 MHz, CDCl₃): δ 8.00 (bs, 1H), 7.45–6.97 (m, 4H), 6.10 (s,1H), 6.08–6.00 (m, 1H), 5.92 (s, 1H), 5.89 (s, 1H), 5.82 (s, 1H), 5.40(dd, J=1.5 Hz, J=17.1 Hz, 1H), 3.38 (bs, 1H), 5.23 (dd, J=1.5 Hz, J=10.5Hz, 1H), 4.42–4.36 (m, 1H), 4.19–4.03 (m, 5H), 3.71 (s, 3H), 3.68–3.17(m, 4H), 2.90 (dd, J=7.8 Hz, J=18.3 Hz, 1H), 2.57 (d, J=18.3 Hz, 1H),2.25 (s, 3H), 2.12 (s, 3H), 2.10 (s, 3H), 1.90 (dd, J=12.3 Hz, J=16.5Hz, 1H), 0.81 (d, J=6.9 Hz, 3H).

¹³C NMR (75 MHz, CDCl₃): δ 178.4, 171.6, 148.6, 146.8, 144.3, 142.7,138.7, 136.2, 133.6, 130.7, 129.8, 126.6, 124.2, 124.1, 120.9, 120.5,117.7, 117.4, 116.7, 112.6, 112.5, 101.0, 74.0, 60.6, 59.0, 57.0, 56.2,56.1, 55.0, 53.3, 41.4, 39.7, 26.3, 24.8, 18.3, 15.5, 9.2.

ESI-MS m/z: Calcd; for C₃₉H₄₄N₆O₆S: 724.8 Found (M+H)⁺: 725.3.

Example 8

To a solution of 20 (270 mg, 0.37 ml) in dioxane (1 ml), 4.2NHCl/dioxane (3.5 ml) was added and the reaction was stirred at 23° C.for 30 min. Then, ethyl acetate (20 ml) and H₂O (20 ml) were added andthe organic layer was decanted. The aqueous phase was basified withsaturated aqueous sodium bicarbonate (60 ml) (pH=8) at 0° C. and then,extracted with CH₂Cl₂ (2×50 ml). The combined organic extracts weredried (sodium sulphate), and concentrated in vacuo. The residue waspurified by flash column chromatography (SiO₂, ethyl acetate:methanol5:1) to afford compound 21 (158 mg, 82%) as a white solid.

Rf: 0.3 (ethyl acetate:methanol 1:1).

¹H NMR (300 MHz, CDCl₃): δ 6.45 (s, 1H), 6.12–6.03 (m, 1H), 5.91 (s,1H), 5.85 (s, 1H), 5.38 (dd, J₁=1.2 Hz, J₂=17.1 Hz, 1H), 5.24 (dd,J₁=1.2 Hz, J₂=10.5 Hz, 1H), 4.23–4.09 (m, 4H), 3.98 (d, J=2.1 Hz, 1H),3.90 (bs, 1H), 3.72 (s, 3H), 3.36–3.02 (m, 5H), 2.72–2.71 (m, 2H), 2.48(d, J=18.0 Hz, 1H), 2.33 (s, 3H), 2.22 (s, 3H), 2.11 (s, 3H), 1.85 (dd,J₁=11.7 Hz, J₂=15.6 Hz, 1H)).

¹³C NMR (75 MHz, CDCl₃): δ 148.4, 146.7, 144.4, 142.8, 138.8, 133.8,130.5, 128.8, 121.5, 120.8, 118.0, 117.5, 116.9, 113.6, 112.2, 101.1,74.3, 60.7, 59.9, 58.8, 56.6, 56.5, 55.3, 44.2, 41.8, 29.7, 26.5, 25.7,15.7, 9.4.

ESI-MS m/z: Calcd. for C₂₉H₃₄N₄O₅: 518.3. Found (M+H)⁺: 519.2.

Example 9

To a solution of 21 (0.64 g, 1.22 ml) in CH₂Cl₂ (6.13 ml), pyridine(0.104 ml 1.28 ml) and 2,2,2-trichloroethyl chloroformate (0.177 ml,1.28 ml) were added at −10° C. The mixture was stirred at thistemperature for 1 h and then, the reaction was quenched by addition of0.1N HCl (10 ml) and extracted with CH₂Cl₂ (2×10 ml). The organic layerwas dried over sodium sulphate and concentrated in vacuo. The residuewas purified by flash column chromatography (SiO₂, (hexane:ethyl acetate1:2) to afford 22 (0.84 g, 98%) as a white foam solid.

Rf: 0.57 (ethyl acetate:methanol 5:1).

¹H NMR (300 MHz, CDCl₃): δ 6.50 (s, 1H), 6.10–6.00 (m, 1H), 6.94 (d,J=1.5 Hz, 1H), 5.87 (d, J=1.5 Hz, 1H), 5.73 (bs, 1H), 5.37 (dq, J=1.5Hz, J₁=17.1 Hz 1H), 5.26 (dq, J₁=1.8 Hz, J₂=10.2 Hz, 1H), 4.60 (d, J=12Hz, 1H), 4.22–4.10 (m, 4H), 4.19 (d, J=12 Hz, 1H), 4.02 (m, 2H), 3.75(s, 3H), 3.37–3.18 (m, 5H), 3.04 (dd, J₁=8.1 Hz, J₂=18 Hz, 1H), 2.63 (d,J=18 Hz, 1H), 2.31 (s, 3H), 2.26 (s, 3H), 2.11 (s, 3H), 1.85 (dd, J=12.3Hz, J₂=15.9 Hz, 1H).

¹³C NMR (75 MHz, CDCl₃) δ 154.3, 148.5, 146.7, 144.5, 142.8, 139.0,133.8, 130.7, 128.7, 121.3, 120.8, 117.8, 117.7, 116.8, 112.7, 101.2,77.2, 74.3, 60.7, 59.9, 57.0, 56.4, 55.3, 43.3, 41.7, 31.6, 26.4, 25.3,22.6, 15.9, 14.1, 9.4.

ESI-MS m/z: Calcd. for C₃₂H₃₅Cl₃N₄O₇: 694.17. Found (M+H)⁺: 695.2.

Example 10

To a solution of 22 (0.32 g, 0.46 ml) in CH₃CN (2.33 ml).diisopropylethylamine (1.62 ml, 9.34 ml), bromomethylmethyl ether (0.57ml, 7.0 ml) and dimethylaminopyridine (6 mg, 0.046 ml) were added at 0°C. The mixture was heated at 30° C. for 10 h. Then, the reaction wasdiluted with dichloromethane (30 ml) and poured in an aqueous solutionof HCl at pH=(10 ml). The organic layer was dried over sodium sulphateand the solvent was eliminated under reduced pressure to give a residuewhich was purified by flash column chromatography (SiO₂, hexane:ethylacetate 2:1) to afford 23 (0.304 g, 88%) as a white foam solid.

Rf: 0.62 (hexane:ethyl acetate 1:3).

¹H NMR (300 MHz, CDCl₃): δ 6.73 (s, 1H), 6.10 (m, 1H), 5.94 (d J=1.5 Hz,1H), 5.88 (d, J=1.5 Hz, 1H), 5.39 (dq, J₁=1.5 Hz, J₂=17.1 Hz, 1H), 5.26(dq, J₁=1.8 Hz, J₂=10.2 Hz, 1H), 5.12 (s, 2H), 4.61 (d, J=12 Hz, 1H),4.55 (t, J=6.6 Hz, 1H), 4.25 (d, J=12 Hz, 1H), 4.22–4.11 (m, 4H), 4.03(m, 2H), 3.72 (s, 3H), 3.58 (s, 3H), 3.38–3.21 (m, 5H), 3.05 (dd, J₁=8.1Hz, J₂=18 Hz, 1H), 2.65 (d, J=18 Hz, 1H), 2.32 (s, 3H), 2.23 (s, 3H),2.12 (s, 3H), 1.79 (dd, J₁=12.3 Hz, J₂=15.9 Hz, 1H);

¹³C NMR (75 MHz, CDCl₃) δ 154.3, 148.6, 148.4, 144.5, 139.0, 133.6,130.6, 130.1, 125.07, 124.7, 124.0, 121.1, 117.7, 112.6, 101.2, 99.2,77.2, 74.4, 74.1, 59.8, 59.8, 57.7, 57.0, 56.8, 56.68, 55.3, 43.2, 41.5,26.4, 25.2, 15.9, 9.3.

ESI-MS m/z: Calcd. for C₃₄H₃₉Cl₃N₄O₈: 738.20. Found (M+H)⁺: 739.0.

Example 11

To a suspension of 23 (0.304 g, 0.41 ml) in 90% aqueous acetic acid (4ml) powder zinc (0.2 g, 6.17 ml) was added and the reaction was stirredfor 7 hour at 23° C. The mixture was filtered through a pad of celitewhich was washed with CH₂Cl₂. The organic layer was washed with anaqueous sat. solution of sodium bicarbonate (pH=9) (15 ml) and driedover sodium sulphate. The solvent was eliminated under reduced pressureto give 24 (0.191 g, 83%) as a white solid.

Rf: 0.3 (ethyl acetate:methanol 5:1).

¹H NMR (300 MHz, CDCl₃): δ 6.68 (s, 1H), 6.09 (m, 1H), 5.90 (d, J=1.5Hz, 1H), 5.83 (d, J=1.5 Hz, 1H), 5.39 (dq, J₁=1.5 Hz, J₂=17.1 Hz, 1H),5.25 (dq, J₁=1.5 Hz, J₂=10.2 Hz, 1H), 5.10 (s, 2H), 4.22–4.09 (m, 3H),3.98 (d, J=2.4 Hz, 1H), 3.89 (m, 1H), 3.69 (s, 3H), 3.57 (s, 3H),3.37–3.17 (m, 3H), 3.07 (dd, J₁=8.1 Hz, J₂=18 Hz, 1H), 2.71 (m, 2H),2.48 (d, J=18 Hz, 1H), 2.33 (s, 3H), 2.19 (s, 3H), 2.17 (s, 3H), 1.80(dd, J₁=12.3 Hz, J₂=15.9 Hz, 1H)

¹³C NMR (75 MHz, CDCl₃): δ 148.5, 148.2, 144.3, 138.7, 133.7, 130.7,129.9, 125.0, 123.9, 121.3, 117.9, 117.5, 113.6, 112.0, 101.0, 99.2,74.0, 59.8, 59.7, 58.8, 57.6, 57.0, 56.2, 55.2, 44.2, 41.5, 31.5, 26.4,25.6, 22.5, 16.7, 14.0, 9.2.

ESI-MS m/z: Calcd. for C₃₁H₃₈N₄O₆: 562.66. Found (M+H)⁺: 563.1.

Example 12

To a solution of 24 (20 mg, 0.035 ml), in H₂O (0.7 ml) and THF (0.7 ml).NaNO₂ (12 mg, 0.17 ml) and 90% aqueous AcOH (0.06 ml) were added at 0°C. and the mixture was stirred at 0° C. for 3 h. After dilution withCH₂Cl₂ (5 ml), the organic layer was washed with water (1 ml), driedover sodium sulphate and concentrated in vacuo. The residue was purifiedby flash column chromatography (SiO₂, hexane:ethyl acetate 2:1) toafford 25 (9.8 mg, 50%) as a white solid.

Rf: 0.34 (hexane:ethyl acetate 1:1).

¹H NMR (300 MHz, CDCl₃): δ 6.71 (s, 1H), 6.11 (m, 1H), 5.92 (d, J=1.5Hz, 1H), 5.87 (d, J=1.5 Hz, 1H), 5.42 (dq, J₁=1.5 Hz, J₂=17.1 Hz, 1H),5.28 (dq, J₁=1.5 Hz., J₂=10.2 Hz, 1H), 5.12 (s, 2H), 4.26–4.09 (m, 3H),4.05 (d, J=2.4 Hz, 1H), 3.97 (t, J=3.0 Hz, 1H), 3.70 (s, 3H), 3.67–3.32(m, 4H), 3.58 (s, 3H), 3.24 (dd, J₁=2.7 Hz, J₂=15.9 Hz, 1H), 3.12 (dd,J₁=8.1 Hz, J₂=18.0 Hz, 1H), 2.51 (d J=18 Hz, 1H), 2.36 (s, 3H), 2.21 (s,3H), 2.12 (s, 3H), 1.83 (dd, J₁=12.3 Hz, J₂=15.9 Hz, 1H)

¹³C NMR (75 MHz, CDCl₃) δ 148.7, 148.4, 138.9, 133.7, 131.1, 129.4,125.1, 123.9, 120.7, 117.6, 117.5, 113.2, 112.3, 101.1, 99.2, 74.0,63.2, 59.8, 59.7, 57.9, 57.7, 57.0, 56.5, 55.2, 41.6, 29.6, 26.1, 25.6,22.6, 15.7, 9.2.

ESI-MS m/z: Calcd. for C₃₁H₃₇N₃O₇: 563.64. Found (M+H)⁺: 564.1.

Example 13

The starting material (2.0 g, 5.90 ml) was added to a suspension ofsodium hydride (354 mg, 8.86 ml) in THF (40 ml) at 23° C., following thesuspension was treated with allyl chloroformate (1.135 ml, 8.25 ml) at23° C. and then refluxed for 3 hours. The suspension was cooled,filtered off, the solid washed with ethyl acetate (100 ml), and thefiltrate was concentrated. The oil crude was ground with hexane (100 ml)and kept at 4° C. overnight. After, the solvent was decanted and thelight yellow slurry was treated with CH₂Cl₂ (20 ml), and precipitatedwith hexane (100 ml). After 10 minutes, the solvent was decanted again.The operation was repeated until appearing a white solid. The whitesolid was filtered off and dried to afford compound 29 (1.80 g, 65%) asa white solid.

¹H-NMR (300 MHz, CDCl₃): δ 7.74 (d, J=7.5 Hz, 2H), 7.62 (d, J=6.9 Hz,2H), 7.33 (t, J=7.5 Hz, 2H), 7.30 (t, J=6.3 Hz, 2H), 5.71 (d, J=7.8 Hz,1H), 4.73 (d, J=7.8 Hz, 2H), 4.59 (m, 1H), 4.11 (t, J=6.0 Hz, 1H), 3.17(dd, J=6.0 Hz, J=2.7 Hz, 2H), 3.20 (dd, J=5.4 Hz, J=2.1 Hz, 2H).

¹³C-NMR (75 MHz, CDCl₃): δ 173.6, 152.7, 144.0, 139.7, 137.8, 126.0,125.6, 123.4, 118.3, 73.4, 52.4, 45.5, 35.8, 33.7.

ESI-MS m/z: Calcd. for C₂₀H₁₈Cl₃NO₄S: 474.8. Found (M+Na)⁺: 497.8

Example 14

A mixture of compound 25 (585 mg, 1.03 ml) and compound 29 (1.47 mg,3.11 ml) were azeotroped with anhydrous toluene (3×10 ml). To a solutionof 25 and 29 in anhydrous CH₂Cl₂ (40 ml) was added DMAP (633 mg, 5.18ml) and EDC.HCl (994 mg, 5.18 ml) at 23° C. The reaction mixture wasstirred at 23° C. for 3 hours. The mixture was partitioned withsaturated aqueous solution of sodium bicarbonate (50 ml) and the layerswere separated. The aqueous layer was washed with CH₂Cl₂ (50 ml). Thecombined organic layers were dried over sodium sulphate, filtered andconcentrated. The crude was purified by flash column chromatography(ethyl acetate/hexane 1:3) to obtain 30 (1.00 g, 95%) as a pale creamyellow solid.

¹H-NMR (300 MHz, CDCl₃): δ 7.72 (m, 2H), 7.52 (m, 2H), 7.38 (m, 2H),7.28 (m, 2H), 6.65 (s, 1H), 6.03 (m, 1H), 5.92 (d, J=1.5 Hz, 1H), 5.79(d, J=1.5 Hz, 1H), 5.39 (m, 1H), 5.29 (dq, J=10.3 Hz, J=1.5 Hz, 1H),5.10 (s, 2H), 4.73 (d, J=11.9 Hz, 1H), 4.66 (d, J=11.9 Hz, 1H), 4.53 (m,1H), 4.36–3.96 (m, 9H), 3.89 (t, J=6.4 Hz, 1H), 3.71 (s, 3H), 3.55 (s,3H), 3.33 (m, 1H), 3.20 (m, 2H), 2.94 (m, 3H), 2.59 (m, 1H), 2.29 (s,3H), 2.23 (s, 3H), 2.02 (s, 3H), 1.83 (dd, J=16.0 Hz, J=11.9 Hz, 1H).

¹³C-NMR (75 MHz, CDCl₃): δ 169.7, 154.0, 148.8, 148.4, 145.7, 144.5,140.9, 139.0, 133.7, 130.9, 130.6, 127.6, 127.0, 124.8, 124.6, 124.1,120.8, 119.9, 118.2, 117.7, 117.3, 112.7, 112.1, 101.3, 99.2, 74.7,73.9, 64.4, 59.8, 57.7, 57.0, 56.8, 55.4, 53.3, 46.7, 41.4, 36.5, 34.7,31.5, 26.4, 24.9, 22.6, 15.7, 14.0, 9.1.

ESI-MS m/z: Calcd. for C₅₁H₅₃Cl₃N₄O₁₀S: 1020.4. Found (M+H)⁺: 1021.2

Example 15

To a solution of 30 (845 mg, 0.82 ml), acetic acid (500 mg, 8.28 ml) and(PPh₃)₂PdCl₂ (29 mg, 0.04 ml) in anhydrous CH₂Cl₂ 20 ml at 23° C. wasadded, dropwise, Bu₃SnH (650 mg, 2.23 ml). The reaction mixture wasstirred at this temperature for 15 min., bubbling was. The crude wasquenched with water (50 ml) and extracted with CH₂Cl₂ (3×50 ml). Theorganic layers were dried over sodium sulphate, filtered andconcentrated. The crude was purified by flash column chromatography(ethyl acetate/hexane in gradient from 1:5 to 1:3) to obtain compound 31(730 mg, 90%) as a pale cream yellow solid.

¹H-NMR (300 MHz, CDCl₃): δ 7.72 (m, 2H), 7.56 (m, 2H), 7.37 (m, 2H),7.30 (m, 2H), 6.65 (s, 1H), 5.89 (s, 1H), 5.77 (s, 1H), 5.74 (s, 1H),5.36 (d, J=5.9 Hz, 1H), 5.32 (d, J=5.9 Hz, 1H), 5.20 (d, J=9.0, 1H),4.75 (d, J=12.0 Hz, 1H), 4.73 (m, 1H), 4.48 (d, J=11.9 Hz, 1H), 4.08 (m,4H), 3.89 (m, 1H), 3.86, (t, J=6.2 Hz, 1H), 3.70 (s, 3H), 3.69 (s, 3H),3.38 (m, 1H), 3.25 (m, 1H), 3.02–2.89 (m, 4H), 2.67 (s, 1H), 2.61 (s,1H), 2.51 (dd, J=14.3 Hz, J=4.5 Hz, 1H), 2.29 (s, 3H), 2.23 (s, 3H),1.95 (s, 3H), 1.83 (m, 1H).

¹³C-NMR (75 MHz, CDCl₃): δ 168.2, 152.5, 148.1, 146.2, 144.4, 144.3,143.3. 139.6, 134.6, 129.7, 129.6, 126.2, 125.6, 123.4, 123.3, 121.6,118.5, 116.3, 110.7, 110.2, 105.1, 99.4, 98.5, 75.2, 73.3, 61.7, 58.4,57.9, 56.3, 56.1, 55.1, 54.7, 53.9, 51.9, 45.2, 40.1, 35.6, 33.3, 24.8,23.3., 14.5, 7.3.

ESI-MS m/z: Calcd. for C₄₈R₄₉Cl₃N₄O₁₀S: 980.3. Found (M+H)⁺: 981.2

Example 16

To a solution of 31 (310 mg, 0.32 ml), in anhydrous CH₂Cl₂ (15 ml) at−10° C. was added a solution of benzeneseleninic anhydride 70% (165 mg,0.32 ml), in anhydrous CH₂Cl₂ (7 ml), via cannula, keeping thetemperature at −10° C. The reaction mixture was stirred at −10° C. for 5min. A saturated solution of sodium bicarbonate (30 ml) was added atthis temperature. The aqueous layer was washed with more CH₂Cl₂ (40 ml).The organic layers were dried over sodium sulphate, filtered andconcentrated. The crude was purified by flash column chromatography(ethyl acetate/hexane in gradient from 1:5 to 1:1) to obtain 32 (287 mg,91%, HPLC: 91.3%) as a pale cream yellow solid and as a mixture of twoisomers (65:35) which were used in the next step.

¹H-NMR (300 MHz, CDCl₃): δ (Mixture of isomers) 7.76 (m, 4H), 7.65 (m,4H), 7.39 (m, 4H), 7.29 (m, 4H), 6.62 (s, 1H), 6.55 (s, 1H), 5.79–5.63(m, 6H), 5.09 (s, 1H), 5.02 (d, J=6.0 Hz, 1H), 4.99 (d, J=6.0 Hz, 1H),4.80–4.63 (m, 6H), 4.60 (m, 1H), 4.50 (m, 1H), 4.38 (d, J=12.8 Hz, J=7.5Hz, 1H), 4.27 (dd, J=12.8 Hz, J=7.5 Hz, 1H), 4.16–3.90 (m, 10H), 3.84(s, 3H), 3.62 (s, 3H), 3.50 (s, 3H), 3.49 (s, 3H), 3.33–2.83 (m, 14H),2.45–2.18 (m, 2H), 2.21 (s, 6H), 2.17 (s, 6H), 1.77 (s, 6H), 1.67 (m,2H).

¹³C-NMR (75 MHz, CDCl₃): δ (Mixture of isomers) 168.6, 168.4, 158.6,154.8, 152.8, 152.5, 147.3, 147.2, 146.8, 144.1, 144.0, 140.8, 139.7,137.1, 129.8, 129.3, 128.4, 128.7, 126.5, 125.5, 123.7, 123.6, 123.5,123.4, 122.2, 121.3, 118.3, 115.8, 115.5, 110.2, 106.9, 103.5, 103.2,100.1, 99.6, 97.9, 97.7, 93.8, 73.4, 70.9, 69.2, 64.9, 62.5, 59.3, 58.9,58.4, 56.7, 56.3, 56.2, 55.4, 55.2, 55.1, 54.9, 54.7, 54.3, 54.1, 53.8,52.8, 45.5, 40.5, 40.0, 39.8, 35.8, 35.5, 33.9, 33.7, 30.1, 28.8, 24.2,24.1, 21.2, 14.5, 14.4, 12.7, 6.0, 5.7.

ESI-MS m/z: Calcd. for C₄₈H₄₉Cl₃N₄O₁₁S: 996.3. Found (M+H)⁺: 997.2

Example 17

The reaction flask was flamed twice, purged vacuum/Argon several timesand kept under Argon atmosphere for the reaction. To a solution of DMSO(39.1 ml, 0.55 ml, 5 equivalents.) in anhydrous CH₂Cl₂ (4.5 ml) wasdropwise added triflic anhydride (37.3 ml, 0.22 ml, 2 equivalents.) at−78° C. The reaction mixture was stirred at −78° C. for 20 minutes, thena solution of 32 (110 mg, 0.11 ml, HPLC: 91.3%) in anhydrous CH₂Cl₂ (1ml for the main addition and 0.5 ml for wash) at −78° C. was added, viacannula. During the addition the temperature was kept at −78° C. in bothflasks and the colour changed from yellow to brown. The reaction mixturewas stirred at −40° C. for 35 minutes. During this period of time thesolution was turned from yellow to dark green. After this time,^(i)Pr₂NEt (153 ml, 0.88 ml, 8 equivalents.) was dropwise added and thereaction mixture was kept at 0° C. for 45 minutes, the colour of thesolution turned to brown during this time. Then t-butanol (41.6 ml, 0.44ml, 4 equivalents.) and 2-′Butyl-1,1,3,3-tetramethylguanidine (132.8 ml,0.77 ml, 7 equivalents.) were dropwise added and the reaction mixturewas stirred at 23° C. for 40 minutes. After this time, acetic anhydride(104.3 ml, 1.10 ml, 10 equivalents.) was dropwise added and-the reactionmixture was kept at 23° C. for 1 hour more. Then the reaction mixturewas diluted with CH₂Cl₂ (20 ml) and washed with aqueous saturatedsolution of NH₄Cl (50 ml), sodium bicarbonate (50 ml), and sodiumchloride (50 ml). The combined organic layers were dried over sodiumsulphate, filtered and concentrated. The residue was purified by flashcolumn chromatography (eluent: ethyl acetate/hexane gradient from 1:3 to1:2) to afford compound 33 (54 mg, 58%) as a pale yellow solid.

¹H-NMR (300 MHz, CDCl₃): δ 6.85 (s, 1H), 6.09 (s, 1H), 5.99 (s, 1H),5.20 (d, J=5.8 Hz, 1H), 5.14 (d, J=5.3 Hz, 1H), 5.03 (m, 1H), 4.82 (d,J=12.2, 1H), 4.63 (d, J=12.0 Hz, 1H), 4.52 (m, 1H), 4.35–4.17 (m, 4H),3.76 (s, 3H), 3.56 (s, 3H), 3.45 (m, 2H), 2.91 (m, 2H), 2.32 (s, 3H),2.28 (s, 3H), 2.21 (s, 3H), 2.12 (m, 2H), 2.03 (s, 3H),

¹³C-NMR (75 MHz, CDCl₃): δ 168.5, 167.2, 152.7, 148.1, 147.1, 144.5,139.6, 139.1, 130.5, 129.0, 123.7, 123.5, 123.3, 118.8, 116.5, 112.1,100.6, 97.8, 73.3, 60.5, 59.4, 59.2, 58.3, 57.6, 57.4, 56.1, 53.3, 53.1,40.6, 40.0, 31.0, 22.2, 18.9, 14.4, 8.1.

ESI-MS m/z: Calcd, for C₃₆H₃₉Cl₃N₄O₁₁S: 842.1. Found (M+H)⁺: 843.1

Example 18

To a solution of 33 (12 mg, 0.014 ml) in dry dichloromethane (1.2 ml)and HPLC grade acetonitrile (1.2 ml) was added at 23° C. sodium iodide(21 mg, 0.14 ml) and freshly distilled (over calcium hydride atatmospheric pressure) trimethylsilyl chloride (15.4 mg, 0.14 ml). Thereaction mixture turned to orange colour. After 15 min the solution wasdiluted with dichloromethane (10 ml) and was washed with a freshlyaqueous saturated solution of Na₂S₂O₄ (3×10 ml). The organic layer wasdried over sodium sulphate, filtered and concentrated. It was obtainedcompound 34 (13 mg, quantitative) as pale yellow solid which was usedwithout further purification.

¹H-NMR (300 MHz, CDCl₃): δ 6.85 (s, 1H), 6.09 (s, 1H), 5.99 (s, 1H),5.27 (d, J=5.8 Hz, 1H), 5.14 (d, J=5.3 Hz, 1H), 5.03 (d, J=11.9 Hz, 1H),4.82 (d, J=12.2, 1H), 4.63 (d, J=13.0 Hz, 1H), 4.52 (m, 1H), 4.34 (m,1H), 4.27 (bs, 1H), 4.18 (m, 2H), 3.76 (s, 3H), 3.56 (s, 3H), 3.44 (m,1H), 3.42 (m, 1H), 2.91 (m, 2H), 2.32 (s, 3H), 2.28 (s, 3H), 2.21 (s,3H), 2.03 (s, 3H).

ESI-MS m/z; Calcd. for C₃₄H₃₅N₄O₁₀S: 798.1. Found (M+H)⁺: 799.1

Example 19

To a solution of 34 (13 mg, 0.016 ml) in a mixture of acetic acid/H₂O(90:10, 1 ml) was added powder Zinc (5.3 mg, 0.081 ml) at 23° C. Thereaction mixture was heated at 70° C. for 6 h. After this time, wascooled to 23° C., diluted with CH₂Cl₂ (20 ml) and washed with aqueoussaturated solution of sodium bicarbonate (15 ml) and aqueous solution ofEt₃N (15 ml). The organic layer was dried over sodium sulphate, filteredand concentrated. The residue was purified by flash columnchromatography with Silica-NH₂ (eluent: ethyl acetate/hexane gradientfrom 0:100 to 50:50) to afford compound 35 (6.8 mg, 77% for two steps)as a pale yellow solid.

¹H-NMR (300 MHz, CDCl₃): δ 6.51 (s, 1H), 6.03 (dd, J=1.3 Hz, J=26.5 Hz,2H), 5.75 (bs, 1H), 5.02 (d, J=11.6 Hz, 1H), 4.52 (m, 1H), 4.25 (m, 2H),4.18 (d, J=2.5 Hz, 1H), 4.12 (dd, J=1.9 Hz, J=11.5 Hz, 1H), 3.77 (s,3H), 3.40 (m, 2H), 3.26 (t, J=6.4 Hz, 1H), 2.88 (m, 2H), 2.30–2.10 (m,2H), 2.30 (s, 3H), 2.28 (s, 3H), 2.18 (s, 3H), 2.02 (s, 3H).

¹³C-NMR (75 MHz, CDCl₃): δ 174.1, 168.4, 147.8, 145.4, 142.9, 140.8,140.1, 131.7, 130.2, 129.1, 128.3, 120.4, 118.3, 117.9, 113.8, 111.7,101.7, 61.2, 59.8, 59.2, 58.9, 54.4, 53.8, 54.4, 41.3, 41.5, 34.1, 23.6,20.3, 15.5, 9.4.

ESI-MS m/z: Calcd. for C₃₁H₃₄N₄O₈S: 622.7. Found (M+H)⁺: 623.2.

Example 20

A solution of N-methylpyridine-4-carboxaldehyde iodide (378 mg, 1.5mmol) in anhydrous DMF (5.8 mL) was treated with anhydrous toluene (2×10mL) to eliminate the amount of water by azeotropic removal of thetoluene. A solution of 35 (134 mg, 0.21 mmol), previously treated withanhydrous toluene (2×10 mL), in anhydrous CH₂Cl₂ (distilled over CaH₂,7.2 mL) was added, via cannula, at 23° C. to this orange solution. Thereaction mixture was stirred at 23° C. for 4 hours. After this time DBU(32.2 μL, 0.21 mmol) was dropwise added at 23° C. and it was stirred for15 minutes at 23° C. A freshly aqueous saturated solution of oxalic acid(5.8 mL) was added to the reaction mixture and was stirred for 30minutes at 23° C. Then the reaction mixture was cooled to 0° C. andNaHCO₃ was portionwise added followed by addittion of aqueous saturatedsolution of NaHCO₃. The mixture was extracted with Et₂O. K₂CO₃ was addedto the aqueous layer and it was extrated with Et₂O. The combined organiclayers were dried over MgSO₄ and the solvent was removed under reducedpressure. The crude was purified by flash column chromatography(AcOEt/hexane from 1/3 to 1/1) to afford compound 36 (77 mg, 57%) aspale yellow solid.

¹H-NMR (300 MHz, CDCl₃): δ 6.48 (s, 1H), 6.11 (d, J=1.3 Hz, 1H), 6.02(d, J=1.3 Hz, 1H), 5.70 (bs, 1H), 5.09 (d, J=11.3 Hz, 1H), 4.66 (bs,1H), 4.39 (m, 1H), 4.27 (d, J=5.6 Hz, 1H), 4.21 (d, J=10.5 Hz, 1H), 4.16(d, J=2.6 Hz, 1H), 3.76 (s, 3H), 3.54 (d, J=5.1 Hz, 1H), 3.42 (d, J=8.5Hz, 1H), 2.88–2.54 (m, 3H), 2.32 (s, 3H), 2.24 (s, 3H), 2.14 (s, 3H),2.04 (s, 3H). ¹³C-NMR (75 MHz, CDCl₃): δ 186.7, 168.5, 160.5, 147.1,146.4, 142.9, 141.6, 140.7, 130.4, 129.8, 121.7 (2C), 120.0, 117.8,117.1, 113.5, 102.2, 61.7, 61.4, 60.3, 59.8, 58.9, 54.6, 41.6, 36.9,29.7, 24.1, 20.3, 15.8, 14.1, 9.6.

ESI-MS m/z: Calcd. for C₃₁H₃₁N₃O₉S: 621.7. Found (M+H)⁺: 622.2.

Example 21

To a solution of 36 (49 mg, 0.08 ml) and2-[3-hydroxy-4-methoxyphenyl]ethylamine (46.2 mg, 0.27 ml) in ethanol(2.5 ml) was added silica gel (105 mg) at 23° C. The reaction mixturewas stirred at 23° C. for 14 h. It was diluted with hexane and pouredinto a column of chromatography (ethyl acetate/hexane from 1/3 to 1/1)to afford Et-770 (55 mg, 90%) as a pale yellow solid.

¹H-NMR (300 MHz, CDCl₃): δ 6.60 (s, 1H), 6.47 (s, 1H), 6.45 (s, 1H),6.05 (s, 1H), 5.98 (s, 1H), 5.02 (d, J=11.4 Hz, 1H), 4.57 (bs, 1H), 4.32(bs, 1H), 4.28 (d, J=5.3 Hz, 1H), 4.18 (d, J=2.5 Hz, 1H), 4.12 (dd,J=2.1 Hz, J=11.5 Hz, 1H), 3.78 (s, 3H), 3.62 (s, 3H), 3.50 (d, J=5.0 Hz,1H), 3.42 (m, 1H), 3.10 (ddd, J=4.0 Hz, J=10.0 Hz, J=11.0 Hz, 1H), 2.94(m, 2H), 2.79 (m, 1H), 2.61 (m, 1H), 2.47 (m, 1H), 2.35 (m, 1H), 2.32(s, 3H), 2.27 (s, 3H), 2.20 (s, 3H), 2.09 (m, 1H), 2.04 (s, 3H).

ESI-MS m/z: Calcd. for C₄₀H₄₂N₄O₁₀S: 770.7. Found (M+H)⁺: 771.2

Example 22

To a solution of 21 (22 mg, 0.042 ml) in CH₂Cl₂ (0.8 ml) was addedphthalic anhydride (6.44 mg, 0.042 ml) and the reaction mixture wasstirred for 2 h at 23° C. Then, carbonyldiimidazole (1 mg, 0.006 ml) wasadded and the mixture was stirred at 23° C. for 7 h. Then,carbonyldiimidazole (5.86 mg, 0.035 ml) was added and the reaction wasstirred at 23° C. for an additional 17 h. The solution was diluted withCH₂Cl₂ (15 ml) and washed with 0.1 N HCl (15 ml). The organic layer wasdried over sodium sulphate, filtered and the solvent was eliminatedunder reduced pressure. The residue was purified by flash columnchromatography (SiO₂, hexane:ethyl acetate 2:1) to afford 27 (26.4 mg,96%) as a white solid.

Rf: 0.58 (ethyl acetate).

¹H NMR (300 MHz, CDCl₃): 7.73–7.64 (m, 4H), 6.40 (s, 1H), 6.12–6.01 (m,1H), 5.63 (s, 1H), 5.58 (d, J=1.5 Hz, 1H), 5.37 (dd, J₁=1.8 Hz, J₂=17.4Hz), 5.23 (dd, J₁=1.8 Hz, J₂=10.5 Hz, 1H), 5.12 (d, J=1.5 Hz, 1H),4.22–4.15 (m, 3H), 4.08 (d, J=1.8 Hz, 1H), 3.68 (s, 3H), 3.59–3.55 (m2H), 3.35 (d, J=8.1 Hz, 1H), 3.27–3.16 (m, 2H), 3.05 (dd, J₁=8.1 Hz,J₂=18.3 Hz, 1H), 2.64 (d, J=18.0 Hz, 1H), 2.30 (s, 3H), 2.24 (s, 3H),2.09 (s, 3H), 1.80 (dd, J₁=11.4 Hz, J₂=15 Hz, 1H);

¹³C NMR (75 MHz, CDCl₃): δ 167.7, 148.9, 146.4, 144.2, 142.6, 139.5,134.0, 133.5, 132.0, 131.0, 128.3, 123.0, 121.3, 120.9, 118.1, 117.5,116.8, 113.6, 112.4, 100.8, 74.5, 60.6, 60.5, 57.7, 56.6, 55.6, 55.5,42.3, 41.7, 26.6, 25.5, 15.9, 9.46.

ESI-MS m/z: Calcd. for C₃₇H₃₅N₄O₇: 648.79. Found (M+H)⁺: 649.3.

Example 23

To a solution of 27 (26 mg, 0.041 ml) in CH₂Cl₂ (11 ml), acetic acid (11ml), (PPh₃)₂PdCl₂ (2.36 mg) and Bu₃SnH (28 ml, 0.10 ml) were added at23° C. After stirring at that temperature for 2 h the reaction waspoured into a pad of flash column (SiO₂, gradient Hex to hexane:ethylacetate 2:1) to afford 28 (24.7 mg, 99%) as a white solid.

Rf: 0.33 (hexane:ethyl acetate 2:1).

¹H NMR (300 MHz, CDCl₃): δ 7.75–7.70 (m, 2H), 7.69–7.65 (m, 2H), 6.39(s, 1H), 5.82 (bs, 1H), 5.50 (d, J=1.5 Hz, 1H), 5.0 (d, J=1.5 Hz, 1H),4.45 (bs, 1H), 4.23–4.19 (m, 2H), 4.10–4.09 (m, 1H), 3.73 (s, 3H),3.60–3.48 (m, 2H), 3.36–3.33 (m, 1H), 3.26–3.20 (m, 1H), 3.14–3.08 (m,1H), 3.98 (d, J=14.4 Hz, 1H), 2.61 (d, J=18.3 Hz, 1H), 2.30 (s, 3H),2.23 (s, 3H), 2.06 (s, 3H), 1.85 (dd, J₁=12 Hz, J₂=15.3 Hz);

¹³C NMR (75 MHz, CDCl₃): δ 167.8, 146.4, 145.1, 143.9, 142.7, 137.1,133.5, 131.9, 130.8, 128.4, 122.9, 120.8, 118.0, 116.8, 114.0, 113.4,106.4, 100.4, 60.6, 60.5, 57.8, 56.6, 55.5, 55.2, 42.6, 41.5, 25.6,25.5, 15.8, 8.9.

ESI-MS m/z: Calcd. for C₃₄H₃₂N₄O₇: 608.6. Found (M+H)⁺: 609.2.

Example 24

To a solution of 28 (357 mg, 0.058 ml) in CH₂Cl₂ (3 ml), acetyl chloride(41.58 ml, 0.58 ml) and pyridine (47.3 ml, 0.58 ml) were added at 0° C.The reaction mixture was stirred for 1 h and then, the solution wasdiluted with CH₂Cl₂ (15 ml) and washed with 0.1 N HCl (15 ml). Theorganic layer was dried over sodium sulphate, filtered, and the solventwas eliminated under reduced pressure. The residue was purified by flashcolumn chromatography (RP-18, CH₃CN:H₂O 60:40) to afford phthalascidin(354 mg, 94%) as a white solid.

Rf: 0.37 (CH₃CN:H₂O 7:3, RP-18).

¹H NMR (300 MHz, CDCl₃): δ 7.72–7.68 (m, 2H), 7.67–7.63 (m, 2H), 6.38(s, 1H), 5.69 (d, J=1.2 Hz, 1H), 5.64 (d, J=1.2 Hz, 1H), 5.30 (bs, 1H),4.25–4.21 (m, 2H), 4.02 (d, J=2.1 Hz, 1H), 3.64–3.62 (m, 5H), 3.33 (d,J=8.4 Hz, 1H), 3.21–3.16 (m, 1H), 3.02 (dd, J₁=8.1 Hz, J₂=18 Hz, 1H),2.76 (dd, J₁=1.8 Hz, J₂=15.6 Hz, 1H), 2.63 (d, J=17.7 Hz, 1H), 2.29 (s,3H), 2.28 (s, 3H), 2.21 (s, 3H), 2.0 (s, 3H), 1.73 (dd, J₁=12.0 Hz,J₂=15.3 Hz, 1H))

¹³C NMR (75 MHz, CDCl₃)): δ 168.5, 167.6, 146.2, 144.2, 142.5, 141.0,140.5, 133.4, 131.8, 130.7, 128.2, 120.9, 120.8, 117.9, 116.4, 113.6,101.1, 60.4, 60.0, 57.0, 56.3, 55.6, 55.4, 41.6, 41.5, 26.5, 25.2, 20.2,15.7, 9.4.

ESI-MS m/z: Calcd. for C₃₆H₃₄N₄O₈: 650. Found (M+H)⁺: 651.2.

Example 25

To a solution of 17 (300 mg, 0.432 ml) in CH₂Cl₂ (2 ml), acetyl chloride(30.7 ml, 0.432 ml) and pyridine (34.9 ml, 0.432 ml) were added at 0° C.The reaction mixture was stirred for 2 h at that temperature and then,the solution was diluted with CH₂Cl₂ (15 ml) and washed with 0.1 N HCl(15 ml). The organic layer was dried over sodium sulphate, filtered, andthe solvent was eliminated under reduced pressure to afford 42 (318 mg,100%) as a white solid that was used in subsequent reactions with nofurther purification.

Rf: 0.5 (ethyl acetate:methanol 5:1).

¹H NMR (300 MHz, CDCl₃), δ 6.66 (s, 1H), 5.93 (d, J=1.2 Hz, 1H), 5.83(d, J=1.2 Hz, 1H), 5.42 (t, J=6.6 Hz, 1H), 5.07 (d, J=5.7 Hz, 1H), 4.98(d, J=5.7 Hz, 1H), 4.16 (d, J=1.8 Hz, 1H), 4.11 (d, J=2.7 Hz, 1H), 3.98(bs, 1H), 3.73–3.61 (m, 2H), 3.64 (s, 3H), 3.52–3.48 (m, 1H), 3.50 (s,3H), 3.33 (d, J=9.6 Hz, 1H), 3.17–3.14 (m, 1H), 2.97–2.87 (m, 1H),2.75–2.70 (d, J=16.8 Hz, 1H), 2.26 (s, 6H), 2.16 (s, 3H), 1.96 (s, 3H),1.70 (dd, J₁=11.7 Hz, J₂=15.6 Hz, 1H), 1.33 (s, 9H), 0.59 (d, J=6.0 Hz,3H).

¹³C NMR (75 MHz, CDCl₃)): δ 172.0, 168.3, 162.3, 148.2, 144.4, 140.4,140.2, 130.9, 130.5, 125.3, 123.4, 120.8, 117.6, 112.7, 111.7, 101.4,99.1, 79.2, 59.5, 58.8, 57.5, 57.4, 56.4, 55.5, 55.0, 41.3, 39.0, 28.2,26.4, 24.6, 19.9, 18.4, 15.4, 9.1.

ESI-MS m/z: Calcd. for C₃₈H₄₉N₅O₁₀: 735.82. Found (M+H)⁺: 736.3.

Example 26

To a solution of 42 (318 mg, 0.432 ml) in CH₂Cl₂ (2.16 ml),trifluoroacetic acid (1.33 ml, 17.30 ml) was added and the reactionmixture was stirred for 3.5 h at 23° C. The reaction was quenched at 0°C. with saturated aqueous sodium bicarbonate (60 ml) and extracted withCH₂Cl₂ (2×70 ml). The combined organic layers were dried (sodiumsulphate) and concentrated in vacuo. The residue was purified by flashcolumn chromatography (SiO₂, ethyl acetate:methanol 20:1) to afford 43(154 mg, 60%) as a white solid.

Rf: 0.22 (ethyl acetate:methanol 5:1).

¹H NMR (300 MHz, CDCl₃), δ 6.47 (s, 1H), 6.22 (bs, 1H), 5.95 (d, J=1.2Hz, 1H), 5.88 (d, J=1.2 Hz, 1H), 4.08–4.06 (m, 2H), 4.01 (bs, 1H), 3.69(s, 3H), 3.49 (d, J=3.6 Hz, 1H), 3.33 (d, J=8.1 Hz, 1H), 3.26–3.22 (m,1H), 2.95 (dd, J₁=8.1 Hz, J₂=18 Hz, 1H), 2.80–2.76 (m, 2H), 2.58 (d,J=18 Hz, 1H), 2.29 (s, 3H), 2.27 (s, 3H), 2.21 (s, 3H), 1.96 (s, 3H),1.77 (dd, J₁=12.3 Hz, J₂=15.6 Hz, 1H), 0.90 (d, J=6.9 Hz, 3H).

¹³C NMR (75 MHz, CDCl₃)): δ 174.8, 169.0, 146.8, 144.4, 142.8, 140.5,140.2, 131.1, 128.8, 120.8, 120.5, 117.1, 112.9, 111.6, 101.5, 60.3,59.0, 56.5, 56.3, 55.6, 55.1, 50.2, 41.6, 39.5, 26.8, 26.3, 24.9, 20.2,15.4, 9.2.

ESI-MS m/z: Calcd. for C₃₁H₃₇N₅O₇: 591.65. Found (M+H)⁺: 592.3.

Example 27

To a solution of 43 (154 mg, 0.26 ml) in CH₂Cl₂ (1.3 ml), phenylisothiocyanate (186 ml, 1.56 ml) was added and the mixture was stirredat 23° C. for 2 h. The reaction was concentrated in vacuo and theresidue was purified by flash column chromatography (SiO₂, gradientHexane to hexane:ethyl acetate 1:1) to afford 44 (120 mg, 63%) as awhite solid.

Rf: 0.41 (ethyl acetate:methanol 5:1).

¹H NMR (300 MHz, CDCl₃), δ 8.17 (s, 1H), 7.49–7.44 (m, 3H), 7.31–7.24(m, 3H), 7.05 (d, J=6.9 Hz, 1H), 5.98 (d, J=1.2 Hz, 1H), 5.87 (d, J=1.2Hz, 1H), 5.52 (bs, 1H), 4.54 (t, J=6.6 Hz, 1H), 4.15 (d, J=2.1 Hz, 1H),4.03 (d, J=2.7 Hz, 2H), 3.80 (bs, 1H), 3.66 (s, 3H), 3.40 (bs, 1H), 3.32(d, J=7.8 Hz, 1H), 3.16 (d, J=11.7 Hz, 1H), 2.82–2.61 (m, 3H), 2.29 (s,3H), 2.20 (s, 3H), 2.01 (s, 3H), 1.99 (s, 3H), 1.80 (dd, J₁=12.0 Hz,J₂=15.9 Hz, 1H), 0.62 (d, J=6.0 Hz, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 178.5, 171.9, 168.7, 146.7, 144.5, 142.6,140.6, 140.3, 136.3, 131.0, 129.9, 128.9, 126.7, 124.4, 120.9, 120.6,117.7, 116.6, 112.7, 111.9, 101.4, 60.4, 58.7, 57.5, 56.1, 55.7, 55.1,53.3, 41.4, 38.8, 26.3, 24.4, 20.2, 18.1, 15.3, 9.2.

ESI-MS m/z: Calcd. for C₃₈H₄₂N₆O₇S: 726.3. Found (M+H)⁺: 727.3.

Example 28

To a solution of 44 (120 mg, 0.165 ml) in dioxane (0.9 ml), 5.3NHCl/dioxane (1.8 ml) was added and the reaction was stirred at 23° C.for 2.5 h. Then, CH₂Cl₂ (10 ml) and H₂O (5 ml) were added to thisreaction and the organic layer was decanted. The aqueous phase wasbasified with saturated aq sodium bicarbonate (20 ml) (pH=8) at 0° C.and then, extracted with CH₂Cl (2×15 ml). The combined organic extractswere dried (sodium sulphate), and concentrated in vacuo to afford 45 (75mg, 87%) as a white solid that was used in subsequent reactions with nofurther purification.

Rf: 0.23 (ethyl acetate:methanol 5:1).

¹H NMR (300 MHz, CDCl₃): δ 6.43 (s, 1H), 5.94 (d, J=1.2 Hz, 1H), 5.87(d, J=1.2 Hz, 1H), 4.10 (d, J=2.1 Hz, 1H), 3.98 (d, J=2.4 Hz, 1H), 3.91(bs, 1H), 3.69 (s, 3H), 3.34–3.25 (m, 2H), 3.05 (dd, J₁=1.8 Hz, J₂=8.1Hz, 1H), 2.80–2.73 (m, 3H), 2.46 (d, J=18 Hz, 1H), 2.30 (s, 3H), 2.28(s, 3H), 2.20 (s, 3H), 1.98 (s, 3H), 1.79 (dd, J₁=12.6 Hz, J₂=16.2 Hz,1H);

¹³C NMR (75 MHz, CDCl₃)): δ 168.7, 146.7, 144.4, 142.9, 140.4, 130.4,128.9, 121.1, 120.8, 117.8, 116.8, 113.6, 111.5, 101.4, 67.6, 60.5,59.8, 58.4, 56.6, 55.8, 55.3, 43.6, 41.8, 31.3, 25.6, 20.2, 15.6, 9.2.

ESI-MS m/z: Calcd. for C₂₈H₃₂N₄O₆: 520.58. Found (M+H)⁺: 521.3.

Example 29

To a solution of 45 (10 mg, 0.02 ml) in CH₂Cl₂ (0.4 ml) was addedphthalic anhydride (2.84 mg, 0.02 ml) and the reaction mixture wasstirred for 2 h at 23° C. Then, carbonyldiimidazole (0.5 mg, 0.003 ml)was added and the mixture was stirred at 23° C. for 7 h. Then,carbonyldiimidazole (2.61 mg, 0.016 ml) was added and the reaction wasstirred at 23° C. for an additional 17 h. The solution was diluted withCH₂Cl₂ (10 ml) and washed with 0.1 N HCl (5 ml). The organic layer wasdried over sodium sulphate, filtered, and the solvent was eliminatedunder reduced pressure. The residue was purified by flash columnchromatography (RP-18, CH₃CN:H₂O 60:40) to afford phthalascidin (11.7mg, 93%) as a white solid.

Rf: 0.37 (CH₃CN:H₂O 7:3, RP-18).

¹H NMR (300 MHz, CDCl₃): δ 7.72–7.68 (m, 2 h), 7.67–7.63 (m, 2 h), 6.38(s, 1H), 5.69 (d, J=1.2 Hz, 1H), 5.64 (d, J=1.2 Hz, 1H), 5.30 (bs, 1H),4.25–4.21 (m, 2 h), 4.02 (d, J=2.1 Hz, 1H), 3.64–3.62 (m, 5H), 3.33 (d,J=8.4 Hz, 1H), 3.21–3.16 (m, 1H), 3.02 (dd, J₁=8.1 Hz, J₂=18 Hz, 1H),2.76 (dd, J₁=1.8 Hz, J₂=15.6 Hz, 1H), 2.63 (d, J=17.7 Hz, 1H), 2.29 (s,3H), 2.28 (s, 3H), 2.21 (s, 3H), 2.0 (s, 3H), 1.73 (dd, J₁=12.0 Hz,J₂=15.3 Hz, 1H));

¹³C NMR (75 MHz, CDCl₃)): δ 168.5, 167.6, 146.2, 144.2, 142.5, 141.0,140.5, 133.4, 131.8, 130.7, 128.2, 120.9, 120.8, 117.9, 116.4, 113.6,101.1, 60.4, 60.0, 57.0, 56.3, 55.6, 55.4, 41.6, 41.5, 26.5, 25.2, 20.2,15.7, 9.4.

ESI-MS m/z: Calcd. for C₃₆H₃₄N₄O₈: 650. Found (M+H)⁺: 651.2.

Example 30

To a solution of 25 (18 mg, 0.032 ml) in DMF (0.05 ml), cat. DMAP (0.5mg, 0.004 ml), imidazole (5 mg, 0.08 ml) and tert-Butyldiphenylsilylchloride (12.5 ml, 0.048 ml) were added at 0° C. and the reactionmixture was stirred for 6 h at 23° C. Water (10 ml) was added at 0° C.and the aqueous phase was extracted with hexane:ethyl acetate 1:10 (2×10ml). The organic layer was dried (sodium sulphate), filtered, and thesolvent was removed under reduced pressure. The crude was purified byflash column chromatography (SiO₂, hexane:ethyl acetate 3:1) to afford26 (27 mg, 88%) as a white solid.

Rf: 0.29 (hexane:ethyl acetate 3:1).

¹H NMR (300 MHz, CDCl₃) δ 7.61–7.58 (m, 2 h), 7.42–7.28 (m, 8H), 6.71(s, 1H), 6.19–6.02 (m, 1H), 5.78 (d, J=1.2 Hz, 1H), 5.64 (d, J=1.2 Hz,1H), 5.40 (dd, J₁=1.2 Hz, J₂=17.1 Hz, 1H), 5.27 (dd, J₁=1.2 Hz, J₂=10.2Hz, 1H), 5.13 (s, 2 h), 4.45 (d, J=2.4 Hz, 1H), 4.24 (d, J=2.1 Hz, 1H),4.17–4.06 (m, 3H), 3.75 (s, 3H), 3.64 (dd, J₁=2.4 Hz, J₂=9.9 Hz, 1H),3.59 (s, 3H), 3.42–3.21 (m, 4H), 3.10 (dd, J₁=8.1 Hz, J₂=17.7 Hz, 1H),2.70 (d, J=17.7 Hz, 1H), 2.33 (s, 3H), 2,26 (s, 3H), 2.11 (s, 3H),2.08–1.89 (m, 1H), 0.87 (s, 9H);

¹³C NMR (75 MHz, CDCl₃): δ 148.5, 148.3, 148.1, 144.0, 139.0, 135.6,135.4, 133.8, 133.1, 132.6, 130.5, 130.3, 129.6, 129.4, 127.5, 127.4,125.1, 124.3, 121.6, 118.5, 117.5, 112.9, 111.7, 100.8, 99.2, 74.0,67.7, 61.5, 59.6, 59.0, 57.7, 57.1, 55.4, 41.6, 29.6, 26.6, 25.5, 18.8,15.8, 9.2.

ESI-MS m/z: Calcd. for C₄₇H₅₅N₃O₇Si: 801.3. Found (M+H)⁺: 802.3.

Example 31

To a solution of 26 (7 mg, 0.0087 ml) in CH₂Cl₂ (0.15 ml), acetic acid(2.5 ml, 0.044 ml), (PPh₃)₂PdCl₂ (0.5 mg, 6.96×10⁻⁴ ml) and Bu₃SnH (3.5ml, 0.013 ml) were added at 23° C. The reaction mixture was stirred atthat temperature for 1 h. The solution was diluted with a mixture ofhexane:ethyl acetate 5:1 (0.5 ml) and poured into a pad of flash column(SiO₂, gradient 5:1 to 1:1 hexane:ethyl acetate) affording ET-11 (5 mg,75%) as a white solid.

Rf: 0.36 (hexane:ethyl acetate 1:5, silica).

¹H NMR (300 MHz, CDCl₃): δ 7.56 (m, 2 h), 7.41–7.25 (m, 8H), 6.67 (s,1H), 5.72 (d, J=1.0 Hz, 1H), 5.58 (d, J=1.0 Hz, 1H), 5.51 (s, 1H), 5.38(d, J=5.75 Hz, 1H), 5.16 (d, J=5.7 Hz, 1H), 4.57 (d, J=2.9 Hz, 1H), 4.21(m, 1H), 4.09 (m, 1H), 3.72 (s, 3H), 3.71 (s, 3H), 3.68 (dd, J₁₌2.1 Hz,J₂₌10.4 Hz, 1H), 3.38–3.26 (m, 3H), 3.11 (dd, J₁₌2.5 Hz, J₂₌15.7 Hz,1H), 3.01 (dd, J₁₌8.9 Hz, J₂₌17.9 Hz, 1H), 2.70 (d, J=17.9 Hz, 1H), 2.31(s, 3H), 2.25 (s, 3H), 2.06 (s, 3H), 1.89 (dd, J₁₌12.1 Hz, J₂₌15.7 Hz,1H), 0.9 (s, 9H).);

¹³C NMR (75 MHz, CDCl₃): δ 149.0, 147.4, 145.3, 144.3, 136.3, 135.7,135.4, 133.2, 130.9, 130.5, 129.6, 129.5, 127.5, 125.0, 118.6, 112.5,112.1, 105.7, 100.5, 99.8, 68.5, 61.5, 59.7, 58.8, 57.7, 56.9, 56.5,55.4, 41.7, 26.6, 26.2, 25.5, 18.9, 15.8, 14.2, 8.7.

ESI-MS m/z: Calcd. for C₄₄H₅₁N₃O₇Si: 761. Found (M+H)⁺: 762.

Example 32

A solution of 2 (3.0 g, 5.46 ml) and phenyl isothiocyanate (3.92 mL,32.76 ml) in CH₂Cl₂ (27 ml) was stirred at 23° C. for 1.5 h. Thereaction mixture was partitioned between CH₂Cl₂ (10 ml) and H₂O (5 ml).The organic layer was dried over sodium sulphate, filtered andconcentrated. The residue was purified by flash column chromatography(SiO₂, gradient Hex to 2:3 hexane:ethyl acetate) to give 3 (3.29 g, 88%)as a yellow solid.

Rf: 0.27 (ACN:H₂O 3:2, RP-C18);

¹H NMR (300 MHz, CDCl₃): δ 7.77 (bs, 1H), 7.42–7.11 (m, 5H), 6.65 (d,1H), 6.29 (s, 1H), 5.6–5.5 (m, 1H), 4.19–4.14 (m, 2 h), 4.08 (d, 1H),3.92 (s, 3H), 3.87–3.65 (m, 6H), 3.77 (s, 3H), 3.37–2.98 (m, 8H), 2.50(d, 1H), 2.31 (s, 3H), 2.20 (s, 3H), 1.96 (d, 1H) 1.87 (s, 3H),1.81–1.75 (m, 1H), 0.96 (d, 3H);

¹³C NMR (75 MHz,

CDCl₃): δ 185.7, 180.9, 178.9, 172.0, 155.7, 147.1, 143.2, 142.4, 136.0,135.1, 130.5, 129.9, 129.3, 128.5, 126.9, 124.4, 120.2, 117.4, 116.3,77.1, 60.9, 58.6, 56.2, 55.8, 55.0, 54.6, 53.5, 41.7, 40.3, 25.1, 24.5,18.4, 15.8, 8.7

ESI-MS m/z: Calcd. for C₃₆H₄₀N₆O₆S: 684.8. Found (M+H)⁺: 685.2.

Example 33

A solution of 3 (0.143 g, 0.208 ml) in 6.5 M HCl/dioxane (150 ml) wasstirred at 23° C. for 6 h. Then, toluene (3 ml) was added to thisreaction and the organic layer was decanted. The residue was partitionedbetween saturated aqueous sodium bicarbonate (3 ml) and CHCl₃ (3×3 ml)The organic layers were dried and concentrated to afford title compoundas a mixture of 4 and 6 (4:6 90:10) which slowly cyclizes to 6 onstanding.

Rf: 0.4 (ethyl acetate:methanol 5:1, silica);

¹H NMR (300 MHz, CDCl₃): δ 6.45 (s, 1H), 4.16 (m, 1H), 4.02 (d, 1H),3.96 (s, 3H), 3.79 (m, 2 h), 3.75 (s, 3H), 3.35 (m, 1H), 3.20–3.00 (m,3H), 2.87 (d, 1H), 2.75 (d, 1H), 2.43 (d, 1H), 2.34 (s, 3H), 2.30 (s,3H), 1.93 (s, 3H), 1.72–1.5 (m, 3H);

ESI-MS m/z: Calcd. for C₂₆H₃₀N₄O₅: 478.5. Found (M+H)⁺: 479.2

Example 34

A solution of 3 (0.143 g, 0.208 ml) in 6.5M HCl/dioxane (150 ml) wasstirred at 23° C. for 1 h. Evaporation of the solvent gave a residuewhich was purified by flash column chromatography (ethylacetate/methanol/triethylamine 100:25:0.1) to give 6 (80 mg, 83%) as ayellow solid.

Rf: 0.26 (ACN:H₂O 3:2, RP-C18);

¹H NMR (500 MHz, CDCl₃): δ 6.46 (s, 1H), 5.9 (bs, 1H) 4.67 (dd, J=18.3Hz, J=7.8 Hz, 1H), 4.24 (d, 1H), 4.16 (s, 3H), 3.93 (d, J=2.7 Hz, 1H),3.8 (m, 2 h), 3.77 (s, 3H), 3.45 (m, 2 h), 3.08 (dd, J=17.9 Hz, J=3.6Hz, 1H), 2.78 (m, 1H), 2.55 (d, 1H), 2.3 (m, 1H), 2.3 (s, 3H) 2.28 (s,3H), 1.90 (s, 3H);

¹³C NMR (75 MHz, CDCl₃): δ 186.2, 162.1, 154.9, 146.9, 145.3, 143.0,130.1, 129.4, 128.1, 125.0, 121.4, 116.4, 116.2, 66.6, 60.7, 60.7, 60.1,59.6, 58.8, 55.6, 54.9, 41.9, 25.3, 24.7, 15.7, 8.9.

ESI-MS m/z: Calcd. for C₂₆H₂₈N₄O₄: 460.5. Found (M+H)⁺: 461.1

Example 35

To a solution of 3 (2.38 g, 3.47 ml) in dioxane (5 ml) 5.3M HCl indioxane (34 ml) was added and the reaction was stirred at 23° C. for 45minutes. Then Ac₂O (51 ml, 539.5 ml) was added and the mixture wasstirred for 4 h. The reaction was cooled at 0° C. and partitionedbetween aqueous saturated Na₂CO₃ (300 ml) and ethyl acetate (300 ml) atthis temperature. The organic phase was dried over sodium sulphate,filtered and concentrated. The residue was purified by flash columnchromatography (SiO₂, gradient CH₂Cl₂ to CH₂Cl₂:ethyl acetate 1:2) togive 5 (1.75 g, 97%) as a yellow solid.

Rf: 0.53 (ACN:H₂O 3:2, RP-C18);

¹H NMR (300 MHz, CDCl₃): δ 6.51 (s, 1H), 5.98 (bs, 1H), 4.84 (dd, 1H),4.17 (d, 1H), 4.00 (d, 1H), 3.99 (s, 3H), 3.85 (bs, 1H), 3.81 (m, 1H),3.74 (s, 3H), 3.70 (d, 1H), 3.23 (m, 1H), 3.11 (dd, 1H), 3.09 (m, 1H),2.93 (m, 2 h), 2.44 (d, 1H), 3.67 (s, 3H), 2.25 (s, 3H), 1.70 (s, 3H),1.60–1.50 (m, 2 h), 1.29 (s, 3H);

¹³C NMR (75 MHz, CDCl₃): δ 185.9, 180.8, 169.9, 160.2, 156.2, 147.0,143.1, 140.4, 136.1. 130.6, 129.6, 127.9, 120.4, 117.2, 61.0, 60.7,58.6, 56.1, 55.7, 55.1, 54.3, 41.8, 41.1, 25.7. 23.9, 22.2, 15.7, 8.7.

ESI-MS m/z: Calcd. for C₂₈H₃₂N₄O₆: 520.6. Found (M+H)⁺: 521.1

Example 36

To a solution of 5 (1.75 g, 3.36 ml) in CH₂Cl₂ (17 ml)diisopropylethylamine (11.71 ml, 67.23 ml), DMAP (20 mg, 0.17 ml) andbromomethyl methyl ether (4.11 ml, 50.42 ml) were added at 0° C. After 6h at 23° C. the reaction was partitioned between CH₂Cl₂ (50 ml) andaqueous saturated sodium bicarbonate (25 ml). The organic layer wasdried over sodium sulphate and the solvent was eliminated under reducedpressure. The crude was purified by flash column chromatography (RP-18,CH₃CN/H₂O 1/1) to give 7 (1.32 g, 70%) as a yellow solid.

Rf: 0.34 (ACN:H₂O 2:3, RP-C18);

¹H NMR (300 MHz, CDCl₃): δ 6.74 (s, 1H), 5.14 (s, 2 h), 4.82 (m, 1H),4.22 (d, 1H), 4.00 (s, 3H), 4.0 (m, 1H), 3.83 (m, 2 h), 3.7 (s, 3H),3.58 (s, 3H), 3.4 (m, 1H), 3.2–2.95 (m, 6H), 2.43 (d, 1H), 2.37 (s, 3H),2.22 (s, 3H), 1.89 (s, 3H), 1.5–1.4 (m, 2 h), 1.31 (s, 3H);

¹³C NMR(75 MHz, CDCl₃): δ 185.9, 180.7, 169.6, 156.2, 148.9, 148.5,140.3, 136.2, 131.3, 130.1, 127.7, 124.6, 123.7, 117.3, 99.5, 99.2,60.9, 59.7, 58.8, 57.7, 56.4, 55.7, 55.0, 54.2, 51.0, 41.6, 41.0, 40.5,25.5, 23.9, 22.3, 19.3, 15.6, 14.6, 8.6.

ESI-MS m/z: Calcd. for C₃₀H₃₆N₄O₇: 564.6. Found (M+H)⁺: 565.3

Example 37

To a solution of 7 (0.37 g, 0.65 ml) in methanol (74 ml) at 0° C. wasadded 1 M sodium hydroxide (130 ml). The reaction was stirred for 15minutes and then quenched at 0° C. with 6M HCl to pH=5. The mixture wasextracted with ethyl acetate (3×50 ml) and the combined organic layerswere dried over sodium sulphate and concentrated in vacuo. The residuewas purified by flash column chromatography (RP-C18 CH₃CN:H₂O 1/:1) toafford 8 (232 mg, 65%) as a yellow oil.

Rf: 0.5 (ACN:H₂O 3:2, RP-C18);

¹H NMR (300 MHz, CDCl₃): δ 6.75 (s, 1H), 5.15 (s, 2 h), 4.86 (m, 1H),4.26 (d, 1H), ), 4.01 (d, 1H), 3.88–3.81 (m, 2 h), 3.70 (s, 3H), 3.58(s, 3H), 3.39 (m, 1H), 3.27–3.21 (m, 1H), 3.18–3.08 (m, 2 h), 3.03–2.97(m, 1H) 2.47 (d, 1H), 2.37 (s, 3H), 2.22 (s, 3H), 1.90 (s, 3H),1.57–1.46 (m, 2 h), 1.33 (s, 3H);

¹³C NMR (75 MHz, CDCl₃): δ 185.3, 180.6, 175.9, 170.1, 151.5, 148.9,148.6, 143.3, 133.7, 131.5, 129.9, 124.7, 123.5, 117.1, 117.0, 99.2,59.8, 58.7, 57.8, 56.3, 55.3, 54.9, 54.3, 41.5, 40.7, 29.6, 25.5, 24.4,22.2, 20.7, 15.7, 8.0.

ESI-MS m/z: Calcd. for C₂₉H₃₄N₄O₇: 550.6. Found (M+H)⁺: 551.2

Example 38

To a degassed solution of compound 8 (240 mg, 0.435 ml) in DMF (30 ml)10% Pd/C (48 mg) was added and the reaction was stirred under H₂(atmospheric pressure.) for 1 h. The reaction was filtered through a padof celite under Argon to a Schlenk tube, as a colourless solution,containing anhydrous Cs₂CO₃ (240 mg, 0.739 ml). Then, bromochloromethane(0.566 ml, 8.71 ml) was added. The tube was sealed and stirred at 90° C.for 3 h. The reaction was cooled and filtrated through celite and washedwith CH₂Cl₂. The organic layer was concentrated and dried (sodiumsulphate) to afford 9 as a brown oil that was used in the next step withno further purification.

Rf: 0.36 (SiO₂, hexane:ethyl acetate 1:5)

¹H NMR (300 MHz, CDCl₃): δ 6.71 (s, 3H), 5.89 (d, 1H), 5.81 (d, 1H),5.63 (bs, 1H), 5.33 (d, 1H), 5.17 (d, 1H), 4.97 (m, 1H), 4.20 (d, 1H),4.09 (m, 1H), 3.99 (m, 1H), 3.68 (m, 1H), 3.65 (s, 6H), 3.59–3.47 (m,4H), 3.37–3.27 (m, 2 h), 3.14–2.97 (m, 2 h), 2.62 (d, 1H), 2.32 (s, 3H),2.20 (s, 3H), 2.08 (s, 3H), 1.72 (m, 1H), 1.36 (s, 3H);

¹³C NMR (75 MHz, CDCl₃): δ 169.8, 149.1, 147.4, 145.5, 136.2, 130.9,130.8, 125.0, 122.9, 117.7, 112.6, 111.8, 106.4, 100.8, 99.8, 59.8,58.9, 57.7, 56.6, 56.4, 55.5, 55.2, 41.6, 40.1, 29.6, 25.9, 25.0, 22.6,15.6, 8.8.

ESI-MS m/z: Calcd. for C₃₀H₃₆SiN₄O₇: 564.6. Found (M+H)⁺: 565.3.

Example 39

To a flask containing 9 (245 mg, 0.435 ml) in DMF, (4 ml), cesiumcarbonate (425 mg, 1.30 ml) and allyl bromide (376 ml, 4.35 ml) wereadded at 0° C. and the mixture was stirred at 23° C. for 1 h. Thereaction was filtered though a pad of celite and partitioned betweenCH₂Cl₂ (25 ml) and H₂O (10 ml). The organic phase was dried (sodiumsulphate) and concentrated at reduced pressure to afford a residue thatwas purified by flash column chromatography (SiO₂, CHCl₃:ethyl acetate1:2) to give 10 as a yellow oil. (113 mg, 43%).

Rf: 0.36 (hexane:ethyl acetate 1:5)

¹H NMR (300 MHz, CDCl₃): δ 6.74 (s, 1H), 6.3–6.0 (m, 1H), 5.94 (d, 1H),5.87 (d, 1H), 5.43–5.36 (m, 2 h), 5.22 (s, 2 h), 5.00 (m, 1H), 4.22 (m,1H), 4.17–4.01 (m, 1H), 3.98 (m, 2 h), 3.71–3.67 (m, 1H), 3.69 (s, 3H),3.62–3.51 (m, 3H), 3.58 (s, 3H), 3.39–3.37 (m, 1H), 3.31–3.26 (m, 3H),3.09 (dd, 1H), 2.56 (d, 1H), 2.36 (s, 3H), 2.21 (s, 3H), 2.11 (s, 3H),2.24–2.10 (m, 1H), 1.82–1.73 (m, 1H), 1.24 (bs, 3H)

¹³C NMR (75 MHz, CDCl₃): δ 169.4, 148.8, 148.3, 139.1, 133.7, 130.9,130.3, 125.2, 120.2, 117.7, 113.1, 112.6, 101.3, 99.3, 74.1, 59.7, 59.3,57.8, 57.0, 56.1, 56.1, 55.2, 41.6, 41.0, 40.9, 29.7, 26.3, 22.5, 15.6,9.3

ESI-MS m/z: Calcd. for C₃₃H₁₀N₄O₇: 604.7. Found (M+H)⁺: 605.3.

Example 40

To a solution of 9 (22 mg, 0.039 ml) in CH₂Cl₂ (0.2 ml), acetyl chloride(2.79 ml, 0.039 ml) and pyridine (3.2 ml, 0.039 ml) were added at 0° C.The reaction mixture was stirred for 1 h and then, the solution wasdiluted with CH₂Cl₂ (10 ml) and washed with 0.1 N HCl (5 ml). Theorganic layer was dried over sodium sulphate, filtered, and the solventwas eliminated under reduced pressure to afford 46 (22 mg, 93%) as awhite solid.

Rf: 0.4 (hexane:ethyl acetate 1:5).

¹H NMR (300 MHz, CDCl₃), δ 6.74 (s, 1H), 5.97 (d, J=0.9 Hz, 1H), 5.91(d, J=0.9 Hz, 1H), 5.12 (d, J=5.7 Hz, 2 h), 5.04 (d, J=5.7 Hz, 1H) 4.90(t, J=6 Hz, 1H), 4.17 (d, J=2.7 Hz, 1H), 4.05 (d, J=2.7 Hz, 1H), 4.01(bs, 1H), 3.71 (s, 3H), 3.57 (s, 3H), 3.50–3.44 (m, 2 h), 3.38–3.36 (m,1H), 3.30–3.26 (m, 1H), 3.00 (dd, J₁=7.8 Hz, J₂=18.0 Hz, 1H) 2.79 (d,J=12.9 Hz, 1H), 2.60 (d, J=18.0 Hz, 1H) 2.35 (s, 3H), 2.32 (s, 3H), 2.21(s, 3H), 2.00 (s, 3H), 1.68 (dd, J₁=11.7 Hz, J₂=15.6 Hz, 1H).

ESI-MS m/z: Calcd. for C_(32h38)N₄O₈: 606.67. Found (M+H)⁺: 607.3.

Example 41

To a solution of 46 (8 mg, 0.013 ml) in dioxane (0.1 ml), 5.3NHCl/dioxane (0.5 ml) was added and the reaction was stirred at 23° C.for 1 h. Then, the solution was diluted with CH₂Cl₂ (5 ml) and washedwith 0.1 N HCl (3 ml). The organic layer was dried over sodium sulphate,filtered, and the solvent was eliminated under reduced pressure toafford 47 (5 mg, 70%) as a white solid.

Rf: 0.4 (hexane:ethyl acetate 1:5).

¹H NMR (300 MHz, CDCl₃), δ 6.51 (s, 1H), 5.97 (d, J=1.2 Hz, 1H), 5.91(d, J=1.2 Hz, 1H), 4.97 (bs, 1H), 4.11 (bs, 1H), 4.04–4.02 (m, 2 h),3.75 (s, 3H),), 3.65 (d, J=2.1 Hz, 2 h), 3.56–3.30 (m, 2 h), 3.04 (dd,J₁=7.5 Hz, J₂=18 Hz, 1H), 2.80 (d, J=14.4 Hz, 1H), 2.59 (d, J=18.3 Hz,1H), 2.33 (s, 3H), 2.24 (s, 3H), 2.00 (s, 3H), 1.76 (dd, J₁=12.0 Hz,J₂=15.9 Hz, 1H), 1.33 (s, 3H), 1.25 (s, 3H).

ESI-MS m/z: Calcd. for C₃₀H₃₄N₄O₇: 562.61. Found (M+H)⁺: 563.3.

Example 42

To a solution of 45 (10 mg, 0.0192 ml) in CH₂Cl₂ (0.3 ml), isovalerylchloride (2.34 ml, 0.0192 ml) and pyridine (1.55 ml, 0.0192 ml) wereadded at 0° C. The reaction mixture was stirred for 1 h and then, thesolution was diluted with CH₂Cl₂ (5 ml) and washed with 0.1 N HCl (3ml). The organic layer was dried over sodium sulphate, filtered, and thesolvent was eliminated under reduced pressure. The residue was purifiedby flash column chromatography (SiO₂, Hex: ethyl acetate 1:2) to afford48 (11 mg, 95%) as a white solid.

Rf: 0.12 (Hex: ethyl acetate 1:2).

¹H NMR (300 MHz, CDCl₃): δ 6.50 (s, 1H), 5.98 (d, J=1.5 Hz, 1H), 5.91(d,J=1.5 Hz, 1H), 5.75 (s, 1H), 5.02 (t, J=5.4 Hz, 1H), 4.10 (d, J=1.5 Hz,1H), 4.06 (d, J=2.7 Hz, 1H), 4.02 (d, J=2.7 Hz, 1H), 3.77 (s, 3H),3.76–3.71 (m, 1H), 3.86–3.28 (m, 3H), 3.04 (dd, J₁=8.1 Hz, J₂=18.3 Hz,1H), 2.78 (d, J=15.9 Hz, 1H), 2.55 (d, J=18 Hz, 1H), 2.32 (s, 6H), 2.26(s, 3H), 1.98 (s, 3H), 1.84–1.68 (m, 2 h), 1.36 (d, J=7.2 Hz, 2 h), 0.69(d, J=6.6 Hz, 3H), 0.62 (d, J=6.6 Hz, 3H).

ESI-MS m/z: Calcd. for C₃₃H₄₀N₄O₇: 604.69. Found (M+H)⁺: 605.3.

Example 43

To a solution of 45 (10 mg, 0.0192 ml) in CH₂Cl₂ (0.3 ml), isovalerylchloride (3.98 ml, 0.0192 ml) and pyridine (1.55 ml, 0.0192 ml) wereadded at 0° C. The reaction mixture was stirred for 1 h and then, thesolution was diluted with CH₂Cl₂ (5 ml) and washed with 0.1 N HCl (3ml). The organic layer was dried over sodium sulphate, filtered, and thesolvent was eliminated under reduced pressure. The residue was purifiedby flash column chromatography (SiO₂, Hex: ethyl acetate 1:2) to afford49 (12.4 mg, 96%) as a white solid.

Rf: 0.7 (ethyl acetate:methanol 10:1).

¹H NMR (300 MHz, CDCl₃): δ 6.50 (s, 1H), 5.98 (d, J=1.5 Hz, 1H), 5.91(d, J=1.5 Hz, 1H), 5.73 (s, 1H), 5.08 (t, J=5.4 Hz, 1H), 4.10 (d, J=1.5Hz, 1H), 4.05 (m., 1H), 4.01 (m, 1H), 3.76 (s, 3H), 3.65–3.61 (m, 1H),3.40–3.27 (m, 3H), 3.03 (dd, J₁=8.1 Hz, J₂=18.6 Hz, 1H), 2.78 (d, J=13.2Hz, 1H), 2.57 (d, J=18.3 Hz, 1H), 2.32 (s, 3H), 2.31 (s, 3H), 2.25 (s,3H), 1.99 (s, 3H), 1.79 (dd, J₁=12.0 Hz, J₂=16.5 Hz, 1H), 1.73–1.42 (m,4H), 1.33–1.18 (m, 10H), 1.03 (m, 2 h), 0.87 (t, J=6.6 Hz, 3H).

ESI-MS m/z: Calcd. for C₃₈H₅₀N₄O₇: 674.83. Found (M+H)⁺: 675.5.

Example 44

To a solution of 45 (14.5 mg, 0.0278 ml) in CH₂Cl₂ (0.3 ml),trans-3-trifluoromethyl cinnamoyl chloride (4.76 ml, 0.0278 ml) andpyridine (2.25 ml, 0.0278 ml) were added at 0° C. The reaction mixturewas stirred for 1 h and then, the solution was diluted with CH₂Cl₂ (5ml) and washed with 0.1 N HCl (3 ml). The organic layer was dried oversodium sulphate, filtered, and the solvent was eliminated under reducedpressure. The residue was purified by flash column chromatography (SiO₂,Hex: ethyl acetate 1:1) to afford 50 (18.7 mg, 94%) as a white solid.

Rf: 0.64 (ethyl acetate:methanol 5:1).

¹H NMR (300 MHz, CH₃OD), δ 7.74–7.55 (m, 4H), 7.23 (d, J=16.0 Hz, 1H),6.34 (s, 1H), 6.12 (d, J=16.0 Hz, 1H), 6.07 (d, J=0.9 Hz, 1H), 5.96 (d,J=0.9 Hz, 1H), 4.39 (d, J=2.4 Hz, 1H), 4.074.05 (m, 1H), 3.81 (bs, 1H),3.46–3.51 (m, 3H), 3.42 (s, 3H), 3.09 (br d, J=12.0 Hz, 1H), 2.94–2.85(m, 2 h), 2.74 (d, J=18.3 Hz, 1H), 2.38 (s, 3H), 2.23 (s, 3H), 2.02 (s,3H), 1.80 (s, 3H), 1.84–1.75 (m, 1H).

¹³C NMR (75 MHz, CDCl₃)): δ 168.7, 165.3, 146.5, 144.7, 142.6, 140.6,138.0, 135.9, 131.0, 130.9, 129.1, 128.6, 125.8, 125.7, 124.5, 124.4,122.7, 121.2, 117.8, 116.5, 113.0, 112.0, 101.7, 60.4, 59.1, 56.5, 56.4,55.6, 55.3, 41.8, 40.3, 26.6, 25.1, 20.3, 15.4, 9.3.

ESI-MS m/z: Calcd. for C₃₉H₃₇F₃N₄O₇: 718.72. Found (M+H)⁺: 719.3.

Example 45

To a solution of 43 (33 mg, 0.0557 ml) in CH₂Cl₂ (0.4 ml), isovalerylchloride (6.79 ml, 0.0557 ml) and pyridine (4.5 ml, 0.0557 ml) wereadded at 0° C. The reaction mixture was stirred for 1 h and then, thesolution was diluted with CH₂Cl₂ (5 ml) and washed with 0.1 N HCl (3ml). The organic layer was dried over sodium sulphate, filtered, and thesolvent was eliminated under reduced pressure. The residue was purifiedby flash column chromatography (SiO₂, Hex: ethyl acetate 1:2) to afford51 (34 mg, 91%) as a white solid.

Rf: 0.09 (Hex: ethyl acetate 1:2).

¹H NMR (300 MHz, CDCl₃): δ 6.46 (s, 1H), 6.10 (bs, 1H), 5.99 (do J=0.9Hz, 1H), 5.90 (d, J=0.9 Hz, 1H), 5.30 (t, J=6.0 Hz, 1H), 4.10–4.05 (m,3H), 3.81 (bs, 1H), 3.74 (s, 3H), 3.54 (bs, 1H), 3.38–3.36 (m, 1H),3.29–3.21 (m, 1H), 3.00 (dd, J₁=8.0 Hz, J₂=18.0 Hz, 1H), 2.25 (s, 3H),2.20 (s, 3H), 2.00 (s, 3H), 1.95–1.90 (m, 3H), 0.87 (d, J=6.6 Hz, 6H),0.76 (d, J=6.0 Hz, 3H).

ESI-MS m/z: Calcd. for C₃₆H₄₅N₅O₈: 675.77. Found (M+H)⁺: 676.3.

Example 46

To a solution of 43 (33 mg, 0.0557 ml) in CH₂Cl₂ (0.4 ml),trans-3-trifluoromethyl cinnamoyl chloride (9.52 ml, 0.0557 ml) andpyridine (4.5 ml, 0.0557 ml) were added at 0° C. The reaction mixturewas stirred for 1 h and then, the solution was diluted with CH₂Cl, (5ml) and washed with 0.1 N HCl (3 ml). The organic layer was dried oversodium sulphate, filtered, and the solvent was eliminated under reducedpressure. The residue was purified by flash column chromatography (SiO₂,Hex: ethyl acetate 1:2) to afford 52 (40 mg, 92%) as a white solid.

Rf: 0.21 (hexane:ethyl acetate 1:2).

¹H NMR (300 MHz, CD₃OD), δ 7.74–7.47 (m, 4H), 6.49 (s, 1H), 6.40 (d,J=15.6 Hz, 1H), 6.00 (d, J=1.5 Hz, 1H), 5.90 (d, J=1.5 Hz, 1H), 5.47 (t,J=6 Hz, 1H), 4.12–4.09 (m, 3H), 3.93 (bs, 1H), 3.71 (s, 3H), 3.59–3.58(m, 1H), 3.38 (d, J=7.8 Hz, 1H), 3.29 (d, J=12.0 Hz, 1H), 3.00 (dd,J₁=8.1 Hz, J₂=18.3 Hz, 1H), 2.79–2.78 (m, 1H), 2.65 (d, J=18.3 Hz, 1H)2.29 (s, 6H), 2.28 (s, 3H), 2.22 (s, 3H), 1.84–1.80 (m, 1H), 0.85–0.84(m, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 171.9, 168.8, 164.4, 146.9, 144.6, 143.0,140.5, 140.5, 139.3, 135.7, 131.1, 131.0, 129.4, 129.1, 126.0, 124.1,124.0, 122.4, 121.1, 120.7, 120.6, 117.7, 116.9, 112.8, 112.0, 101.6,60.6, 59.3, 57.1, 56.3, 55.9, 55.2, 49.0, 41.7, 49.9, 26.5, 25.1, 20.2,18.4, 15.7, 9.3.

ESI-MS m/z: Calcd. for C₄₁H₄₂F₃N₅O₈: 789.8. Found (M+H)⁺: 790.3.

Example 47

To a solution of 43 (10 mg, 0.0169 ml) in CH₂Cl₂ (0.2 ml)trifluoroacetic anhydride (2.38 μl, 0.0169 ml) was added at 23° C. Thereaction mixture was stirred for 5 h and then, the solution was dilutedwith CH₂Cl₂ (5 ml) and washed with 0.1 N HCl (3 ml). The organic layerwas dried over sodium sulphate, filtered, and the solvent was eliminatedunder reduced pressure. The residue was purified by flash columnchromatography (SiO₂, Hex: ethyl acetate 3:2) to afford 53 (10.7 mg,93%) as a white solid.

Rf: 0.57 (ethyl acetate:methanol 5:1).

¹H NMR (300 MHz, CDCl₃) δ 6.45 (s, 1H), 6.00 (d, J=1.2 Hz, 1H), 5.90 (d,J=1.2 Hz, 1H), 5.87 (bs, 1H), 5.32 (bs, 1H), 4.12(d, J=2.1 Hz, 1H), 4.08(d, J=1.8 Hz, 1H), 3.78–3.56 (m, 3H), 3.72 (s, 3H), 3.40 (d, J=8.1 Hz,1H), 3.25 (d, J=9.3 Hz, 1H), 3.00 (dd, J₁=8.4 Hz, J₂=18.0 Hz, 1H), 2.77(dd, J₁=2.1 Hz, J₂=15.9 Hz, 1H), 2.68 (d, J=18.6 Hz, 1H), 2.30 (s, 3H),2.28 (s, 3H), 2.22 (s, 3H), 2.00 (s, 3H), 1.75 (dd, J₁=11.4 Hz, J₂=15.9Hz, 1H), 0.69 (d, J=6.3 Hz, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 170.1, 168.6, 156.0, 147.0, 144.6, 143.0,140.6, 140.4, 131.0, 129.4, 120.9, 120.7, 117.6, 116.8, 112.4, 112.1,101.6, 60.5, 59.0, 57.1, 56.3, 55.6, 55.2, 48.7, 41.6, 39.4, 26.5, 24.9,20.2, 17.8, 15.4, 9.2.

ESI-MS m/z: Calcd. for C₃₃H₃₆F₃N₅O₈: 687.63. Found (M+H)⁺: 688.66.

Example 48

To a solution of 19 (11 mg, 0.0169 ml) in CH₂Cl₂ (0.2 ml)trifluoroacetic anhydride (2.38 ml, 0.0169 ml) was added at 23° C. Thereaction mixture was stirred for 5 h and then, the solution was dilutedwith CH₂Cl₂ (5 ml) and washed with 0.1 N HCl (3 ml). The organic layerwas dried over sodium sulphate, filtered, and the solvent was eliminatedunder reduced pressure. The residue was purified by flash columnchromatography (SiO₂, Hex: ethyl acetate 3:2) to afford 54 (10.7 mg,93%) as a white solid.

Rf: 0.6 (ethyl acetate:methanol 5:1).

¹H NMR (300 MHz, CDCl₃) δ 7.33 (d, J=6.3 Hz, 1H), 6.45 (s, 1H), 6.04 (m,1H), 5.95 (d, J=1.5 Hz, 1H), 5.84 (d, J=1.5 Hz, 1H), 5.32 (m, 2 h), 5.21(m, 1H), 4.11 (m, 4H), 3.73 (s, 3H), 3.64 (m, 2 h), 3.51 (m, 1H), 3.37(d, J=7.8 Hz, 1H), 3.22 (m, 2 h), 3.03 (dd, 1H, J₁=8.1 Hz, J₂=18.3 Hz,1H), 2.60 (d, J=18.3 Hz, 1H), 2.29 (s, 3H), 2.24 (s, 3H), 2.08 (s, 3H),1.86 (dd, J₁=12 Hz, J₂=16.2 Hz, 1H), 0.82 (d, J=7.2 Hz, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 170.0, 156.0, 148.4, 147.1, 144.3, 143.0,138.7, 133.8, 130.5, 129.4, 120.6, 120.4, 117.6, 117.5, 117.0, 113.5,112.5, 112.4, 101.1, 74.1, 66.8, 60.4, 59.3, 56.9, 56.6, 56.3, 55.4,48.7, 41.6, 40.1, 26.2, 25.0, 17.6, 15.4, 9.1.

ESI-MS m/z: Calcd. for C₃₅H₃₉F₃N₅O₇: 685.69. Found (M+H)⁺: 686.3.

Example 49

To a solution of 54 (100 mg, 0.415 ml) in CH₂Cl₂ (4 ml), acetic acid (40ml), (PPh₃)₂PdCl₂ (8.4 mg, 0.012 ml) and Bu₃SnH (157 ml, 0.56 ml) wereadded at 23° C. After stirring at that temperature for 2 h the reactionwas poured into a pad of flash column (SiO₂, gradient Hex tohexane:ethyl acetate 2:1) to afford 55 (90 mg, 96%) as a white solid.

Rf: 0.6 (hexane:ethyl acetate 1:2).

¹H NMR (300 MHz, CDCl₃) δ 7.55 (d, J=7.2 Hz, 1H), 6.45 (s, 1H), 5.90 (d,J=1.2 Hz, 1H), 5.82 (d, J=1.2 Hz, 1H), 5.37 (t, J=6.0 Hz, 1H), 4.15 (d,J=2.1 Hz, 1H), 4.04 (d, J=1.8 Hz, 1H), 3.70 (s, 3H), 3.66–3.53 (m, 2 h),3.37–3.31 (m, 2 h), 3.19–3.15 (d, J=11.7 Hz, 1H), 3.08–3.00 (m, 2 h),2.56 (d, J=18.3 Hz, 1H), 2.30 (s, 3H), 2.24 (s, 3H), 2.04 (s, 3H), 1.91(dd, J₁=12.0 Hz, J₂=15.6 Hz, 1H), 0.84 (d, J=6.9 Hz, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 170.1, 156.3, 147.3, 144.9, 144.4, 143.3,136.7, 130.7, 129.3, 120.6, 117.6, 117.4, 114.4, 112.1, 107.7, 101.0,85.8, 60.5, 59.3, 56.5, 56.4, 56.2, 55.2, 48.9, 41.6, 40.9, 25.7, 25.3,18.0, 15.6, 8.7.

ESI-MS m/z: Calcd. for C_(32h35)F₃N₅O₇: 645.63. Found (M+H)⁺: 646.2.

Example 50

To a solution of 17 (200 mg, 0.288 ml) in CH₂Cl₂ (1.44 ml),trifluoroacetic acid (888 ml, 11.53 ml) was added and the reactionmixture was stirred for 4 h at 23° C. The reaction was quenched at 0° C.with saturated aqueous sodium bicarbonate (60 ml) and extracted withethyl acetate (2×70 ml). The combined organic layers were dried (sodiumsulphate) and concentrated in vacuo to afford 56 (147 mg, 93%) as awhite solid that was used in subsequent reactions with no furtherpurification.

Rf: 0.19 (ethyl acetate:methanol 5:1).

¹H NMR (300 MHz, CD₃OD), 66.48 (s, 1H), 5.88, d, J=0.9 Hz, 1H), 5.81 (d,J=0.9 Hz, 1H), 4.35 (d, J=2.4 Hz, 1H), 4.15 (d, J=1.8 Hz, 1H), 3.99–3.98(m, 1H), 3.70 (s, 3H), 3.52–2.96 (m, 7H), 2.68 (d, J=18.3 Hz, 1H), 2.24(s, 3H), 2.23 (s, 3H), 2.06 (s, 3H), 1.85 (dd, J₁=11.7 Hz, J₂=15.6 Hz,1H), 0.91 (d, J=6.6 Hz, 3H).

¹³C NMR (75 MHz, CD₃OD): δ 173.2, 149.1, 145.6, 144.9, 138.0, 132.2,130.6, 121.4, 119.6, 117.4, 114.3, 109.2, 102.5, 82.3, 60.4, 58.4, 58.3,57.8, 56.6, 50.1, 42.3, 41.6, 27.8, 26.2, 19.5, 15.5, 9.8.

ESI-MS m/z: Calcd. for C₂₉H₃₅N₅O₆: 549.62. Found (M+H)⁺: 550.3.

Example 51

To a solution of 56 (10 mg, 0.018 ml) in CH₂Cl₂ (0.4 ml), phenylisothiocyanate (13 ml, 0.109 ml) was added and the reaction was stirredat 23° C. for 1.5 h. The mixture was concentrated in vacuo and theresidue was purified by flash column chromatography (SiO₂, gradientHexane to 1:1 hexane:ethyl acetate) to afford 57 (8 mg, 65%) as a whitesolid.

Rf: 0.57 (ethyl acetate:methanol 10:1).

¹H NMR (300 MHz, CDCl₃): δ 7.88 (bs, 1H), 7.41–7.36 (m, 2 h), 7.27–7.22(m, 1H), 7.02–7.00 (d, J=7.8 Hz, 2 h), 6.71 (d, J=7.2 Hz, 1H), 6.31 (s,1H), 6.17 (bs, 1H), 5.93 (d, J=1.2 Hz, 1H), 5.83 (d, J=1.2 Hz, 1H), 5.55(bs, 1H), 5.20–5.17 (m, 1H), 4.16 (d, J=1.8 Hz, 1H), 4.05 (bs, 1H), 4.02(d, J=2.4 Hz, 1H), 3.79 (s, 3H), 3.75–3.71 (m, 1H), 3.35 (d, J=7.8 Hz,1H), 3.28–3.19 (m, 2 h), 3.12–2.97 (m, 2 h), 2.50 (d, J=18.3 Hz, 1H),2.32 (s, 3H), 2.21 (s, 3H), 2.15–2.09 (dd, J₁=11.4 Hz, J₂=15.9 Hz, 1H),1.95 (s, 3H), 0.88 (d, J=6.9 Hz, 3H).

¹³C NMR (75 MHz, CDCl₃): δ 178.5, 171.7, 147.2, 145.0, 144.3, 143.3,137.0, 135.7, 130.6, 130.4, 129.6, 127.5, 124.3, 120.6, 117.7, 117.2,115.3, 112.1, 108.3, 100.9, 60.9, 59.5, 56.7, 56.5, 56.2, 55.2, 54.1,41.7, 41.1, 26.3, 25.4, 18.5, 15.8, 9.0.

ESI-MS m/z: Calcd. for C₃₆H₄₀N₆O₆S: 684.81. Found (M+H)⁺: 685.3.

Example 52

To a solution of 57 (45 mg, 0.065 ml) in CH₂Cl₂ (0.5 ml), acetylchloride (4.67 ml, 0.065 ml) and pyridine (5.3 ml, 0.065 ml) were addedat 0° C. The reaction mixture was stirred for 3 h and then, the solutionwas diluted with CH₂Cl₂ (10 ml) and washed with 0.1 N HCl (5 ml). Theorganic layer was dried over sodium sulphate, filtered, and the solventwas eliminated under reduced pressure. The residue was purified by flashcolumn chromatography (RP-18, CH₃CN:H₂O 40:60) to afford 58 (14 mg, 28%)as a white solid.

Rf: 0.34 (CH₃CN:H₂O 7:15).

¹H NMR (300 MHz, CDCl₃), δ 11.90 (d, J=6.6 Hz, 1H), 7.45–7.40 (m, 3H),7.18–7.15 (m, 2 h), 6.58 (s, 1H), 6.00 (d, J=1.2 Hz, 1H), 5.89 (d, J=1.2Hz, 1H), 5.70 (s, 1H), 5.37 (t, J=4.8 Hz, 1H), 4.48 (m, 1H), 4.23 (bs,1H), 4.07 (bs, 2 h), 3.85–3.75 (m, 1H), 3.70 (s, 3H), 3.46–3.41 (m, 2h), 3.24–3.20 (m, 1H), 3.00–2.95 (m, 1H), 2.87–2.75 (m, 1H), 2.31 (s,3H), 2.28 (s, 3H), 2.24 (s, 3H), 2.00 (s, 3H), 1.85 (dd, J₁=11.4 Hz,J₂=15.6 Hz, 1H), 1.66 (s, 3H), 0.82 (d, J=6.0 Hz, 3H).

¹³C NMR (75 MHz, CDCl₃)): δ 182.6, 174.3, 171.0, 146.6, 144.6, 142.7,142.3, 140.7, 140.2, 131.3, 129.8, 129.3, 128.9, 128.8, 121.5, 120.4,117.3, 116.6, 112.8, 112.0, 111.3, 101.5, 60.5, 59.0, 57.6, 56.2, 55.9,55.3, 55.1, 41.6, 39.4, 27.8, 26.5, 24.8, 20.2, 17.1, 15.5, 9.3.

ESI-MS m/z: Calcd. for C₄₀H₄₄N₆O₈S: 768.88. Found (M+H)⁺: 769.2.

Example 53

A solution of 57 (130 mg, 0.189 ml) in dioxane (1 ml), 5.3N HCl/dioxane(1.87 ml) was added and the reaction was stirred at 23° C. for 4 h.Then, CH₂Cl₂ (15 ml) and H₂O (10 ml) were added to this reaction and theorganic layer was decanted. The aqueous phase was basified withsaturated aq sodium bicarbonate (60 ml) (pH=8) at 0° C. and then,extracted with ethyl acetate (2×50 ml). The combined organic extractswere dried (sodium sulphate), and concentrated in vacuo to afford 59 (63mg, 70%) as a white solid.

Rf: 0.15 (ethyl acetate:methanol 5:1).

¹H NMR (300 MHz, CDCl₃), δ 6.67 (s, 1H), 5.99 (d, J=0.9 Hz, 1H), 5.91(d, J=1.2 Hz, 1H), 5.10 (bs, 1H), 4.32 (d, J=7.2 Hz, 1H), 4.25 (dd,J₁=3.6 Hz, J₂=9.3 Hz, 1H), 3.7 (s, 3H), 3.71–3.64 (m, 2 h), 3.50 (dd,J₁=2.4 Hz, J₂=15.9 Hz, 1H), 3.42–3.37 (m, 2 h), 3.16 (dd, J₁=3.6 Hz,J₂=12.9 Hz, 1H), 2.57 (dd, J₁=9.3 Hz, J₂=12.9 Hz, 1H), 2.27 (s, 3H),2.11 (s, 3H), 1.91 (dd, J₁=12.0 Hz, J₂=15.9 Hz, 1H).

ESI-MS m/z: Calcd. for C₂₆H₃₀N₄O₅: 478.5. Found (M+H)⁺: 479.3.

Example 54

A solution of 43 (20 mg, 0.0338 mmol) in CH₂Cl₂ (0.3 ml), cinnamoylchloride (5.63 mg, 0.0338 mmol) and pyridine (2.73 ml, 0.0338 mmol) wereadded at 0° C. The reaction mixture was stirred for 1 h and then, thesolution was diluted with CH₂Cl₂ (10 ml) and washed with 0.1 N HCl (5ml). The organic layer was dried over sodium sulphate, filtered, and thesolvent was eliminated under reduced pressure. The residue was purifiedby flash column chromatography (SiO₂, EtOAc:MeOH 20:1) to afford 60 (22mg, 90%) as a white solid.

Rf: 0.56 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃), 7.51 (s, 1H), 7.50–7.47 (m, 2H), 7.36–7.35 (m,2H), 6.43 (s, 1H), 6.36 (brd, J=15.9 Hz, 2H), 6.01 (d, J=1.5 Hz, 1H),5.90 (brd, J=1.5 Hz, 2H), 5.42 (t, J=6.0 Hz 1H), 4.12–4.07 (m, 3H),3.96–3.95 (m, 1H), 3.73 (bs, 3H), 3.58 (bs, 2H), 3.39 (d, J=8.7 Hz, 1H),3.25 (d, J=11.7 Hz, 1H), 3.0 (dd, J₁=7.5 Hz, J₂=17.7 Hz, 1H), 2.78 (d,J=15.9 Hz, 1H), 2.67 (d, J=16.5 Hz, 1H), 2.29 (s, 6H), 2.23 (s, 3H),1.99 (s, 3H), 1.82 (dd, J₁=11.4 Hz, J₂=15.6 Hz, 1H), 0.83 (d, J=6.0 Hz,3H).

¹³C NMR (75 MHz, CDCl₃)): δ. 172.0, 165.0, 146.9, 144.6, 143.1, 141.0,140.5, 134.8, 131.0, 129.7, 129.1, 128.8, 127.8, 125.5, 123.8, 123.0,121.1, 120.5, 117.7, 116.9, 112.8, 112.0, 101.9, 60.6, 59.2, 57.1, 56.4,55.9, 55.3, 48.8, 41.7, 40.0, 26.5, 25.1, 20.3, 18.5, 15.7, 9.3.

ESI-MS m/z: Calcd. for C₄₀H₄₃N₅O₈: 721.8. Found (M+H)⁺: 722.3.

Example 55

A solution of 45 (19 mg, 0.0364 mmol) in CH₂Cl₂ (0.3 ml),heptafluorobutyryl chloride (5.44 ml, 0.0364 mmol) and pyridine (2.95ml, 0.0364 mmol) were added at 0° C. The reaction mixture was stirredfor 1 h and then, the solution was diluted with CH₂Cl₂ (10 ml) andwashed with 0.1 N HCl (5 ml). The organic layer was dried over sodiumsulphate, filtered, and the solvent was eliminated under reducedpressure. The residue was purified by flash column chromatography (SiO₂,EtOAc:MeOH 20:1) to afford 61 (11.7 mg, 45%) as a white solid.

Rf: 0.76 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 6.46 (s, 1H), 6.12 (bs, 1H), 5.98 (d, J=1.2Hz, 1H), 5.93 (d, J=1.2 Hz, 1H), 5.72 (bs, 1H), 4.13–4.11 (m, 2H), 4.0(d, J=2.4 Hz, 1H), 3.98–3.96 (m, 1H), 3.73 (s, 3H), 3.39 (d, J=7.5 Hz,1H), 3.39–3.28 (m, 2H), 3.09 (dd, J₁=8.1 Hz, J₂=18.0 Hz, 1H), 2.80 (d,J=16.2 Hz, 1H), 2.46 (d, J=18.3 Hz, 1H), 2.32 (s, 6H), 2.21 (s, 3H),1.99 (s, 3H), 1.80 (dd, J₁=12.0 Hz, J₂=16.2 Hz, 1H).

ESI-MS m/z: Calcd. for C₃₂H₃₁F₇N₄O₇: 716.6. Found (M+H)⁺: 717.2.

Example 56

A solution of 43 (24 mg, 0.04 mmol) in CH₂Cl₂ (0.3 ml), butyryl chloride(4.15 ml, 0.04 mmol) and pyridine (3.28 ml, 0.04 mmol) were added at 0°C. The reaction mixture was stirred for 1 h and then, the solution wasdiluted with CH₂Cl₂ (10 ml) and washed with 0.1 N HCl (5 ml). Theorganic layer was dried over sodium sulphate, filtered, and the solventwas eliminated under reduced pressure. The residue was purified by flashcolumn chromatography (SiO₂, EtOAc:MeOH 20:1) to afford 62 (24 mg, 90%)as a white solid.

Rf: 0.35 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 6.47 (s, 1H), 6.10 (d, J=6.5 Hz, 1H), 6.0 (d,J=1.5 Hz, 1H), 5.91 (d, J=1.5 Hz, 1H), 5.86 (bs, 1H), 5.31 (d, J=6.9 Hz,1H), 4.11–4.06 (m, 3H), 3.85–3.81 (m, 1H), 3.75 (s, 3H), 3.59–3.53 (m,2H), 3.38 (d, J=7.5 Hz, 1H), 3.27–3.22 (m, 1H), 3.0 (dd, J₁=7.8 Hz,J₂=17.4 Hz, 1H), 2.79 (d, J=15.3 Hz, 1H), 2.63 (d, J=17.7 Hz, 1H), 2.31(s, 3H), 2.0 (s, 3H), 1.80 (dd, J₁=12.0 Hz, J₂=15.9 Hz, 1H), 1.58 q,J=7.2 Hz, 2H), 0.89 (t, J=7.2 Hz, 3H), 0.76 (d, J=6.6 Hz, 3H).

ESI-MS m/z: Calcd. for C₃₅H₄₃N₅O₈: 661.64. Found (M+H)⁺: 662.3

Example 57

A solution of 43 (19 mg, 0.0364 mmol) in CH₂Cl₂ (0.3 ml), cinnamoylchloride (6.06 mg, 0.0364 mmol) and pyridine (2.95 ml, 0.0364 mmol) wereadded at 0° C. The reaction mixture was stirred for 1 h and then, thesolution was diluted with CH₂Cl₂ (10 ml) and washed with 0.1 N HCl (5ml). The organic layer was dried over sodium sulphate, filtered, and thesolvent was eliminated under reduced pressure. The residue was purifiedby flash column chromatography (SiO₂, EtOAc:MeOH 20:1) to afford 63(20.1 mg, 85%) as a white solid.

Rf: 0.65 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 7.39–7.29 (m, 5H), 6.42, (s, 1H), 6.01 (d,J=1.5 Hz, 1H), 5.92 (d, J=1.5 Hz, 1H), 5.73 (bs, 1H), 5.24 (t, J=6.8 Hz,1H), 4.12–4.08 (m, 3H), 3.66–3.64 (m, 2H), 3.58 (bs, 3H), 3.36 (d, J=8.7Hz, 1H), 3.29 (d, J=12.0 Hz, 1H), 2.98 (dd, J₁=8.1 Hz, J₂=18 Hz, 1H),2.33 (s, 6H), 2.29 (s, 3H), 2.01 (s, 3H), 1.84 (dd, J₁=12.0 Hz, J₂=15.9Hz, 1H). ).

ESI-MS m/z: Calcd. for C₃₇H₃₈N₄O₇: 650.72. Found (M+H)⁺: 651.2.

Example 58

A solution of 43 (20 mg, 0.0338 mmol) in CH₂Cl₂ (0.3 ml),3-chloropropionyl chloride (3.22 ml, 0.0338 mmol) and pyridine (2.73 ml,0.0338 mmol) were added at 0° C. The reaction mixture was stirred for 1h and then, the solution was diluted with CH₂Cl₂ (10 ml) and washed with0.1 N HCl (5 ml). The organic layer was dried over sodium sulphate,filtered, and the solvent was eliminated under reduced pressure. Theresidue was purified by flash column chromatography (SiO₂, EtOAc:MeOH20:1) to afford 64 (20.5 mg, 89%) as a white solid.

Rf: 0.32 (EtOAc:Hexane 5:1).

¹H NMR (300 MHz, CDCl₃) 6.48 (s, 3H), 6.28 (m, 1H), 5.99 (d., J=1.2 Hz,1H) 5.91 (d, J=1.2 Hz, 1H), 5.86 (bs, 1H), 5.31 (m, 1H), 4.08–4.07 (m,3H), 3.75 (s, 3H), 3.72–3.53 (m, 5H), 3.39 (d, J=8.1 Hz, 1H), 3.24 (d,J=12.0 Hz, 1H), 3.00 (dd, J₁=8.1 Hz, J₂=18.0 Hz, 1H), 2.79 (d, J=13.5Hz, 1H), 2.50 (t, J=6.3 Hz, 2H), 2.32 (s, 3H), 2.28 (s, 3H), 2.25 (s,3H), 2.0 (s, 3H), 1.79 (dd, J₁=12.3 Hz, J₂=14.8 Hz, 1H), 0.81 (d, J=6.3Hz, 3H).

Example 59

A solution of 43 (19 mg, 0.0364 mmol) in CH₂Cl₂ (0.3 ml), butyrylchloride (3.78 ml 0.0364 mmol) and pyridine (2.95 ml, 0.0364 mmol) wereadded at 0° C. The reaction mixture was stirred for 1 h and then, thesolution was diluted with CH₂Cl₂ (10 ml) and washed with 0.1 N HCl (5ml). The organic layer was dried over sodium sulphate, filtered, and thesolvent was eliminated under reduced pressure. The residue was purifiedby flash column chromatography (SiO₂, EtOAc:MeOH 20:1) to afford 64 (19mg, 87%) as a white solid.

Rf: 0.60 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) 6.50 (s, 1H), 5.98 (d, J=1.5 Hz, 1H), 5.91 (d,J=1.5 Hz, 1H), 5.75 (s, 1H), 5.01 (t, J=6.4 Hz, 1H), 4.10–4.09 (m, 1H),4.06 (d, J=2.1 Hz, 1H), 4.03–4.02 (m, 1H), 3.76 (s, 3H), 3.67–3.60 (m,1H), 3.42–3.35 (m, 2H), 3.29 (d, J=12.0 Hz, 1H), 3.02 (dd, J₁=7.8 Hz,J₂=17.7 Hz, 1H), 2.79 (d, J=14.1 Hz, 1H), 2.56 (d, J=18.3 Hz, 1H), 2.32(s, 3H), 2.31 (s, 3H), 2.25 (s, 3H), 1.78 (dd, J₁=12.0 Hz, J₂=15.9 Hz,1H), 1.63 (s, 3H), 1.53–1.46 (m, 2H), 1.28–1.16 (m, 2H), 0.68 (t J=7.2Hz, 3H).

ESI-MS m/z: Calcd. for C₃₂H₃₈N₄O₇: 590.67. Found (M+H)⁺: 591.2.

Example 60

To a solution of 50 (31.7 mg, 0.044 mmol) in CH₃CN/H₂O (1.5 ml/0.5 ml),AgNO₃ (225 mg, 1.32 mmol) was added and the reaction was stirred at 23°C. for 17 h. Then brine (10 ml) and Aq sat NaHCO₃ (10 ml) were added at0° C. and the mixture was stirred for 15 min, filtered through a pad ofcelite and washed with CH₂Cl₂ (20 ml). The solution was decanted and theorganic layer was dried and concentrated in vacuo. The residue waspurified by flash column chromatography (SiO₂, EtOAc:MeOH 5:1) to afford66 (16 mg, 51%) as a white solid.

Rf: 0.26 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 7.66–7.42 (m, 4H), 7.20 (bs 1H), 6.44 (s, 1H),5.97 (b. J=1.2 Hz, 1H), 5.90 (d, J=1.2 Hz, 1H), 5.76 (bs, 1H), 5.28 (bs,1H), 4.54 (bs, 1H), 4.43 (bs, 1H), 4.00 (bs, 1H), 3.68–3.57 (m, 4H),3.47 (d, J=3.3 Hz, 1H), 3.40 (d, J=11.7 Hz, 1H), 3.17 (d. J=6.9 Hz, 1H),2.92 (dd, J₁=8.1 Hz, J₂=17.7 Hz, 1H), 2.74 (d, J=17.1 Hz, 1H) 2.48 (d,J=18.6 Hz, 1H), 2.32 (s, 6H), 2.28 (s, 3H), 1.99 (s, 3H), 1.76 (dd,J₁=12.0 Hz, J₂=16.2 Hz, 1H).

ESI-MS m/z: Calcd. for C₃₇H₃₈F₃N₃O₈: 709. Found (M⁺−17): 692.3.

Example 61

To a solution of 53 (57 mg, 0.0828 mmol) in CH₃CN/H₂O (1.5 mL/0.5 ml),AgNO₃ (650 mg, 3.81 mmol) was added and the reaction was stirred at 23°C. for 24 h. Then, brine (10 ml) and Aq sat NaHCO₃ (10 ml) were added at0° C. and the mixture was stirred for 15 min, filtered through a pad ofcelite and washed with CH₂Cl₂ (20 ml). The solution was decanted and theorganic layer was dried and concentrated in vacuo. The residue waspurified by flash column chromatography (SiO₂, EtOAc:MeOH 5:1) to afford67 (28 mg, 50%) as a white solid.

Rf: 0.28 (EtOAc:MeOH 10:1).

¹H NMR (300 MHz, CDCl₃) d 6.47 (s, 1H), 5.97 (s, 1H), 5.88 (s, 1H), 5.35(bs, 1H), 4.51 (bs, 1H), 4.41 (bs, 1H), 4.12–4.05 (m, 1H), 4.00 (d,J=2.7 Hz, 1H), 3.77 (s, 3H), 3.64 (bs, 1H), 3.46 (d, J=3.3 Hz, 1H), 3.34(d, J=11.4 Hz, 1H), 3.18 (d, J=7.5 Hz, 1H), 2.95 (dd, J₁=8.4 Hz, J₂=18.3Hz, 1H), 2.70 (d, J=15.6 Hz, 1H), 2.48 (d, J=17.7 Hz, 1H), 2.28 (s, 3H),2.27 (s, 3H), 2.26 (s 3H), 1.98 (s, 3H), 1.68 (dd, J₁=12 Hz, J₂=15.6 Hz,1H), 0.86 (d, J=6.3 Hz, 3H).

ESI-MS m/z: Calcd. for C₃₂H₃₇F₃N₄O₉: 678.66. Found (M⁺−17): 661.2.

Example 62

To a solution of 48 (32 mg, 0.0529 mmol) in CH₃CN/H₂O (1.5 ml/0.5 ml),AgNO₃ (270 mg, 1.58 mmol) was added and the reaction was stirred at 23°C. for 24 h. Then, brine (10 ml) and Aq sat NaHCO₃ (10 ml) were added at0° C. and the mixture was stirred for 15 min, filtered through a pad ofcelite and washed with CH₂Cl₂ (20 ml). The solution was decanted and theorganic layer was dried and concentrated in vacuo. The residue waspurified by flash column chromatography (SiO₂, EtOAc:MeOH 5:1) to afford68 (18 mg, 56%) as a white solid.

Rf: 0.40 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) d 6.50 (s, 1H), 5.95 (d, J=1.2 Hz, 1H), 5.88 (d,J=1.2 Hz, 1H), 5.23 (d, J=6.9 Hz, 1H), 4.45 (d, J=3.3 Hz, 1H), 4.38 (s,1H), 4.01 (d, J=2.4 Hz, 1H), 3.78 (m, 1H), 3.77 (s, 3H), 3.41–3.37 (m,1H), 3.17–3.15 (m, 1H), 2.96 (dd, J₁=7.8 Hz, J₂=18.0 Hz, 1H), 2.70 (d,J=15.3 Hz, 1H), 2.40 (d, J=18.0 Hz, 1H), 2.30 (s, 6H), 2.27 (s, 3H),1.76–1.65 (m, 1H), 1.35–1.25 (m, 2H), 0.89–0.82 (m, 1H), 0.69 (d, J=6.6Hz, 3H), 0.58 (d, J=6.6 Hz, 3H)

Example 63

To a solution of 51 (27 mg, 0.04 mmol) in CH₃CN/H₂O (1.5 ml/0.5 ml),AgNO₃ (204 mg, 1.19 mmol) was added and the reaction was stirred at 23°C. for 24 h. Then, brine (10 ml) and Aq sat NaHCO₃ (10 ml) were added at0° C. and the mixture was stirred for 15 min, filtered through a pad ofcelite and washed with CH₂Cl₂ (20 ml). The solution was decanted and theorganic layer was dried and concentrated in vacuo. The residue waspurified by flash column chromatography (SiO₂, EtOAc:MeOH 5:1) to afford69 (10 mg, 38%) as a white solid.

Rf: 0.38 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) d 6.48 s, 1H), 6.16 (bs, 1H), 5.98 (d, J=1.5 Hz,1H), 5.89 (d, J=1.5 Hz, 1H), 5.33 (t, J=6.0 Hz, 1H), 4.50 (m, 1H), 4.40(m, 1H), 4.11–4.09 (m, 1H), 4.00 (d, J=2.6 Hz, 1H), 3.78 (s, 3H),3.41–3.32 (m, 3H), 3.18 (d, J=8.4 Hz, 1H), 2.94 (dd, J₁=8.4 Hz, J₂=18.3Hz, 1H), 2.70 (d, J=14.4 Hz, 1H), 4.45 (d, J=18.3 Hz, 1H), 2.31 (s, 3H),2.28 (s, 3H), 2.27 (s, 3H), 2.04 (s, 3H), 2.00–1.86 (m, 3H), 1.73 (m,1H), 0.87 (d, J=6.3 Hz, 6H).

Example 64

To a solution of 63 (15 mg, 0.023 mmol) in CH₃CN/H₂O (1.5 ml/0.5 ml).AgNO₃ (118 mg, 0.691 mmol) was added and the reaction was stirred at 23°C. for 24 h. Then, brine (10 ml) and Aq sat NaHCO₃ (10 ml) were added at0° C. and the mixture was stirred for 15 min, filtered through a pad ofcelite and washed with CH₂Cl₂ (20 ml). The solution was decanted and theorganic layer was dried and concentrated in vacuo. The residue waspurified by flash column chromatography (SiO₂, EtOAc:MeOH 5:1) to afford70 (20.1 mg, 85%) as a white solid.

Rf: 0.43 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) d 7.38–7.28 (m, 5H), 6.48 (s, 1H), 5.98 (d,J=1.5 Hz, 1H), 5.91 (d, J=1.5 Hz, 1H), 5.75 (bs, 1H), 5.38 (brd, 1H),5.30 (bs, 1H), 4.53 (in, 1H), 4.42 (in, 1H), 4.02 (d, J=2.7 Hz, 1H),3.78–3.65 (m, 5H), 3.46–3.40 (m, 2H), 3.17 (d, J=7.8 Hz, 1H), 2.94 (dd,J₁=7.8 Hz, J₂=17.7 Hz, 1H), 2.73 (d, J=16.8 Hz, 1H), 2.45 d, J=18.0 Hz,1H), 2.31 (s, 6H), 2.28 (s, 3H), 1.97 (s, 3H), 1.77 (dd, J₁=12.0 Hz,J₂=15.3 Hz, 1H).

Example 65

To a solution of 65 (25 mg, 0.042 mmol) in CH₃CN/H₂O (1.5 ml/0.5 ml),AgNO₃ (215.56 mg, 1.269 mmol) was added and the reaction was stirred at23° C. for 24 h. Then, brine (10 ml) and Aq sat NaHCO₃ (10 ml) wereadded at 0° C. and the mixture was stirred for 15 min, filtered througha pad of celite and washed with CH₂Cl₂ (20 ml). The solution wasdecanted and the organic layer was dried and concentrated in vacuo. Theresidue was purified by flash column chromatography (SiO₂, EtOAc:MeOH5:2) to afford 71 (16 mg 65%) as a white solid.

Rf: 0.0.5 (EtOAc:MeOH 5:2).

¹H NMR (300 MHz, CDCl₃) d 6.50 (s, 1H), 5.95 (d, J=1.5 Hz, 1H), 5.78 (s,1H), 5.19 (bs, 1H), 4.45 (d, J=3.3 Hz, 1H), 4.37 (bs, 1H), 4.11 (brd,J=4.8 Hz, 1H), 4.01 (d, J=2.1 Hz, 1H), 3.76 (s, 1H), 3.71–3.69 (m, 1H),3.49–3.35 (m, 1H), 3.24 (d, J=13.5 Hz, 1H), 3.15 (d, J=9.3 Hz, 1H), 2.95(dd, J₁=8.1 Hz, J₂=17.7 Hz, 1H), 2.70 (d J=15.6 Hz, 1H), 2.40 (d, J=18.0Hz, 1H), 2.31 (s, 3H), 2.29 (s, 3H), 2.26 (s, 3H), 1.96 (s, 3H),1.75–1.66 (m, 1H), 1.52–1.17 (m, 2H), 0.66 (t, J=7.2 Hz, 3H).

Example 66

To a solution of 45 (35 mg, 0.0672 mmol) in CH₂Cl₂ (0.3 mL),hydrocinnamoyl chloride (11.58 μl, 0.0672 mmol) and pyridine (5.43 μL,0.0672 mmol) were added at 0° C. The reaction mixture was stirred for1.5 h and then, the solution was diluted with CH₂Cl₂ (10 mL) and washedwith 0.1 N HCl (5 mL). The organic layer was dried over Na₂SO₄,filtered, and the solvent was eliminated under reduced pressure. Theresidue was purified by flash column chromatography (SiO₂, gradient Hex:ethyl acetate 2:1 to ethyl acetate) to afford 72 (30 mg, 68%) as a whitesolid.

Rf: 0.51 (ethyl acetate:MeOH 10:1).

¹H NMR (300 MHz, CDCl₃) δ 7.23–7.12 (m, 3H), 7.05–7.00 (m, 2H), 5.97 (d,J=1.2 Hz, 1H), 5.91 (d, J=1.2 Hz, 1H), 5.73 (s, 1H), 5.04 (brt, 1H),4.08 (d, J=2.4 Hz, 1H), 4.02 (bs, 1H), 4.00 (d, J=2.4 Hz, 1H), 3.58 (dd,J₁=4.5 Hz, J₂=13.8 Hz, 1H), 3.47 (bs, 3H), 3.33 (d, J=7.5 Hz, 1H), 3.29(dt, J₁=2.7 Hz, J₂=11.7 Hz, 1H), 3.00 (dd, J₁=7.8 Hz, J₂=18.3 Hz, 1H),2.79 (d, J=14.1 Hz, 1H), 2.58–2.50 (m, 3H), 2.32 (s, 3H), 2.29 (s, 3H),2.03 (s, 3H), 2.01 (s, 3H), 1.94–1.76 (m, 4H).

ESI-MS m/z: Calcd. for C₃₇H₄₀N₄O₇: 652.7. Found (M+Na)⁺: 675.3.

Example 67

To a solution of 45 (45 mg, 0.0576 mmol) in CH₂Cl₂ (0.3 mL), phenylacetyl chloride (7.61 μl, 0.0576 mmol) and pyridine (4.6 μL, 0.0576mmol) were added at 0° C. The reaction mixture was stirred for 1 h andthen, the solution was diluted with CH₂Cl₂ (10 mL) and washed with 0.1 NHCl (5 mL). The organic layer was dried over Na₂SO₄, filtered, and thesolvent was eliminated under reduced pressure. The residue was purifiedby flash column chromatography (SiO₂, gradient Hex:ethyl acetate 3:1 toHex:ethyl acetate 1:1) to afford 73 (25.8 mg, 70%) as a white solid.

Rf: 0.5 (Hex:ethyl acetate:MeOH 5:10:2).

¹H NMR (300 MHz, CDCl₃) δ 7.18–7.17 (m, 3H), 6.85 (bs, 2H), 6.54 (s,1H), 5.89 (d, J=1.5 Hz, 1H), 5.83 (d, J=1.5 Hz, 1H), 5.76 (s, 1H), 5.08(bs, 1H), 4.12 (d, J=2.1 Hz, 1H), 4.09 (d, J=2.1 Hz, 1H), 3.98 (bs, 1H),3.73 (s, 3H), 3.51–3.46 (m, 2H), 3.35 (d, J=8.4 Hz, 11H), 3.25 (dt,J₁=2.7 Hz, J₂=12.0 Hz, 1H), 3.03 (d, J=8.7 Hz, 1H), 3.02–2.94 (m, 2H),2.75 (d, J=16.8 Hz, 1H), 2.63 (d, J=18.0 Hz, 1H), 2.35 (s, 3H), 2.30 (s,3H), 2.22 (s, 3H9, 1.98 (s, 3H), 1.80 (dd, J₁=12.0 Hz, J₂=16.2 Hz, 1H).

ESI-MS m/z: Calcd. for C₃₆H₃₈N₄O₇: 638.7. Found (M+1)⁺: 639.2.

Example 68

To a solution of 45 (30 mg, 0.0576 mmol) in CH₂Cl₂ (0.3 mL), propyonylchloride (5 μL, 0.0576 mmol) and pyridine (4.6 μL, 0.0576 mmol) wereadded at 0° C. The reaction mixture was stirred for 1 h and then, thesolution was diluted with CH₂Cl₂ (10 mL) and washed with 0.1 N HCl (5mL). The organic layer was dried over Na₂SO₄, filtered, and the solventwas eliminated under reduced pressure. The residue was purified by flashcolumn chromatography (SiO₂, gradient Hex:ethyl acetate 5:1 to Hex:ethylacetate 1:1 to ethyl acetate) to afford 74 (23 mg, 70%) as a whitesolid.

Rf: 0.59 ((Hex:ethyl acetate:MeOH 5:10:2).

¹H NMR (300 MHz, CDCl₃) δ 6.50 (s, 1H), 5.97 (d, J=1.2 Hz, 1H), 5.91 (d,J=1.2 Hz, 1H), 5.76 (s, 1H), 5.00 (t, 1H), 4.09 (d, J=1.2 Hz, 1H), 4.04(bs, 2H), 3.74 (s, 3H), 3.62 (dd, J₁=6.6 Hz, J₂=13.2 Hz, 1H), 3.43 (bs,1H), 3.37 (d, J=8.4 Hz, 1H), 3.29 (d, J=12.0 Hz, 1H), 3.02 (dd, J₁=8.1Hz, J₂=218.3 Hz, 1H), 2.80 (d, J=14.4 Hz, 1H), 2.55 (d, J=18.0 Hz, 1H),2.31 (s, 3H), 2.24 (s, 3H), 2.00 (s, 3H), 1.78 (dd, J₁=12.0 Hz, J₂=15.6Hz, 1H), 1.64–1.50 (m, 2H), 0.70 (t, J=7.8 Hz, 3H).

ESI-MS m/z: Calcd. for C₃₁H₃₆N₄O₇: 576.6. Found (M+1)⁺: 577.2.

Example 69

To a solution of 45 (15 mg, 0.0288 mmol) in CH₂Cl₂ (0.25 mL), myristoylchloride (7.83 μL, 0.0288 mmol) and pyridine (2.3 μL, 0.0288 mmol) wereadded at 0° C. The reaction mixture was stirred for 1 h and then, thesolution was diluted with CH₂Cl₂ (10 mL) and washed with 0.1 N HCl (5mL). The organic layer was dried over Na₂SO₄, filtered, and the solventwas eliminated under reduced pressure. The residue was purified by flashcolumn chromatography (SiO₂, gradient Hex:ethyl acetate 6:1 to Hex:ethylacetate 1:1) to afford 75 (15 mg, 71%) as a white solid.

Rf: 0.65 (Hex:ethy acetate:MeOH 10:10:1).

¹H NMR (300 MHz, CDCl₃) δ 6.49 (s, 1H), 5.97 (d, J=1.2 Hz, 1H), 5.91 (d,J=1.2 Hz, 1H), 5.72 (s, 1H), 4.99 (t, 1H), 4.09 (d, J=1.5 Hz, 1H), 4.05(d, J=1.5 Hz, 1H), 4.02 (bs, 1H), 3.76 (s, 3H), 3.61–3.59 (m, 1H), 3.39(bs, 1H), 3.35 (d, J=7.8 Hz, 1H), 3.29 (d, J=12.3 Hz, 1H), 3.04 (dd,J₁=8.1 Hz, J₂=18.3 Hz, 1H), 2.78 (d, J=15.6 Hz, 1H), 2.55 (d, J=18.3 Hz,1H), 2.32 (s, 6H), 2.25 (s, 3H), 1.99 (s, 3H), 1.78 (dd, J₁=12.3 Hz,J₂=15.0 Hz, 1H), 1.25–1.24 (m, 12H), 0.87 (d, J=6.0 Hz, 3H).

ESI-MS m/z: Calcd. for C₄₂H₅₈N₄O₇: 730.9. Found (M+1)⁺: 731.4.

Example 70

To a solution of 45 (15 mg, 0.0288 mmol) in CH₂Cl₂ (0.25 mL), stearoylchloride (9.7 μL, 0.0288 mmol) and pyridine (2.3 μL, 0.0288 mmol) wereadded at 0° C. The reaction mixture was stirred for 1 h and then, thesolution was diluted with CH₂Cl₂ (10 mL) and washed with 0.1 N HCl (5mL). The organic layer was dried over Na₂SO₄, filtered, and the solventwas eliminated under reduced pressure. The residue was purified by flashcolumn chromatography (SiO₂, gradient Hex:ethyl acetate 3:1 to Hex:ethylacetate 1:1) to afford 76 (16 mg, 70%) as a white solid.

Rf: 0.46 (Hex:ethyl acetate:MeOH 10:10:1).

¹H NMR NMR (300 MHz, CDCl₃) δ 6.49 (s, 1H), 5.98 (d, J=1.5 Hz, 1H), 5.91(d, J=1.5 Hz, 1H), 5.73 (s, 1H), 4.99 (t, J=5.7 Hz, 1H), 4.09 (d, J=1.8Hz, 1H), 4.05 (d, J=2.4 Hz, 1H), 4.01 (bs, 1H), 3.76 (s, 3H), 3.61–3.59(m, 1H), 3.38 (bs, 1H), 3.36 (d, J=7.2 Hz, 1H), 3.28 (d, J=12.0 Hz, 1H),3.03 (dd, J₁=7.8 Hz, J₂=18.3 Hz, 1H), 2.78 (d, J=15.9 Hz, 1H), 2.57 (d,J=18.3 Hz, 1H), 2.32 (s, 3H), 2.31 (s, 3H), 2.24 (s, 3H), 1.99 (s, 3H),1.77 (dd, J₁=11.7 Hz, J₂=15.6 Hz, 1H), 1.25–1.24 (m, 16H), 0.87 (d,J=6.3 Hz, 3H).

ESI-MS m/z: Calcd. for C₄₆H₆₆N₄O₇: 786.4. Found (M+22)⁺: 809.5.

Example 71

To a solution of 45 (31 mg, 0.0595 mmol) in CH₂Cl₂ (0.3 mL), hexanoylchloride (8.32 μL, 0.0595 mmol) and pyridine (4.8 μL, 0.0595 mmol) wereadded at 0° C. The reaction mixture was stirred for 1.5 h and then, thesolution was diluted with CH₂Cl₂ (10 mL) and washed with 0.1 N HCl (5mL). The organic layer was dried over Na₂SO₄, filtered, and the solventwas eliminated under reduced pressure. The residue was purified by flashcolumn chromatography (SiO₂, gradient Hex:ethyl acetate 3:2 to ethylacetate) to afford 77 (26 mg, 70%) as a white solid.

Rf: 0.65 (ethyl acetate MeOH 10:1).

¹H NMR (300 MHz, CDCl₃) δ 6.50 (s, 1H), 5.98 (d, J=1.5 Hz, 1H), 5.91 (d,J=1.5 Hz, 1H), 5.74 (s, 1H), 5.00 (t, J=5.4 Hz, 1H), 4.09 (d, J=2.7 Hz,1H), 4.05 (d, J=2.4 Hz, 1H), 4.01 (bs, 1H), 3.76 (s, 3H), 3.61–3.58 (m,1H), 3.02 (dd, J₁=8.1 Hz, J₂=18.3 Hz, 1H), 2.78 (d, J=14.4 Hz, 1H), 2.56(d, J=18.3 Hz, 1H), 2.31 (s, 6H), 2.25 (s, 3H), 2.00 (s, 3H), 1.78 (dd,J₁=12.0 Hz, J₂=15.9 Hz, 1H), 1.53–1.40 (m, 2H), 1.29–1.12 (m, 4H),1.07–0.97 (m, 2H), 0.81 (t, J=7.5 Hz, 3H).

ESI-MS m/z: Calcd. for C₃₄H₄₂N₄O₇: 618.7. Found (M+1)⁺: 619.3.

Example 72

To a solution of 45 (20 mg, 0.0384 mmol) in CH₂Cl₂ (0.3 mL),trans-crotonyl chloride (3.68 μL, 0.0384 mmol) and pyridine (3.1 μL,0.0384 mmol) were added at 0° C. The reaction mixture was stirred for 1h and then, the solution was diluted with CH₂Cl₂ (10 mL) and washed with0.1 N HCl (5 mL). The organic layer was dried over Na₂SO₄, filtered, andthe solvent was eliminated under reduced pressure. The residue waspurified by flash column chromatography (SiO₂, gradient Hex:ethylacetate 4:1 to ethyl acetate) to afford 78 (16 mg, 71%) as a whitesolid.

Rf: 0.55 (ethyl acetate:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 6.50–6.40 (m, 1H), 6.46 (s, 1H), 5.97 (d,J=1.5 Hz, 1H), 5.91 (d, J=1.5 Hz, 1H), 5.77 (s, 1H), 5.08 (bst, 1H),4.10 (d, J=1.5 Hz, 1H), 4.05 (m, 2H), 3.78 (s, 3H), 3.67 (bs, 1H),3.42–3.29 (m, 3H), 3.04 (dd, J₁=8.1 Hz, J₂=18.3 Hz, 1H), 2.78 (d, J=15.3Hz, 1H), 2.53 (d, J=18.3 Hz, 1H), 2.32 (s, 3H), 2.26 (s, 3H), 1.98 (s,3H), 1.79 (dd, J₁=12.0 Hz, J₂=15.6 Hz, 1H), 1.70 (dd, J₁=1.2 Hz, J₂=6.6Hz, 3H).

ESI-MS m/z: Calcd. for C₃₂H₃₆N₄O₇: 588.6. Found (M+1)⁺: 589.3.

Example 73

To a solution of 45 (50 mg, 0.096 mmol) in CH₂Cl₂ (0.5 mL). Cbz-L-Val-OH(24.12 mg, 0.096 mmol) and carbonyl diimidazole (18.7 mg, 0.115 mmol)were added at 0° C. The reaction mixture was stirred for 16 h at roomtemperature and then, the solution was diluted with CH₂Cl₂ (15 mL) andwashed with 0.1 N HCl (10 mL). The organic layer was dried over Na₂SO₄,filtered, and the solvent was eliminated under reduced pressure. Theresidue was purified by flash column chromatography (SiO₂, Hex:EtOAc4:1) to afford 79 (25 mg, 34%) as a white solid.

Rf: 0.7 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 7.33–7.28 (m, 5H), 6.45 (s, 1H), 5.96 (s, 1H),5.90 (bs, 1H), 5.82 (s, 1H), 5.53 (bs, 1H), 5.09 (bs, 1H), 5.05 (d,J=3.3 Hz, 2H), 4.16 (bs, 1H), 4.09 (d, J=2.4 Hz, 1H), 4.02 (bs, 1H),3.75 (s, 31H), 3.74 (m, 1H), 3.37–3.35 (m, 2H), 3.26–3.21 (m, 3H), 3.00(dd, J₁=8.1 Hz, J₂=18.3 Hz, 1H), 2.77 (d, J=15.6 Hz, 1H), 2.55 (d,J=18.0 Hz, 1H), 2.30 (s, 3H), 2.27 (s, 3H), 2.25 (s, 3H), 1.98 (s, 3H),1.70–1.66 (m, 1H), 0.65 (d, J=6.6 Hz, 3H).

ESI-MS m/z: Calcd. for C₄₁H₄₇N₅O₉: 753.8. Found (M+1)⁺: 754.2.

Example 74

To a solution of 72 (18 mg, 0.0275 mmol) in CH₃CN/H₂O (1.5 mL/0.5 mL).AgNO₃ (140.5 mg, 0.827 mmol) was added and the reaction was stirred at23° C. for 24 h. Then, brine (10 mL) and Aq sat NaHCO₃ (10 mL) wereadded at 0° C. and the mixture was stirred for 15 min, filtered througha pad of celite and washed with CH₂Cl₂ (20 mL). The solution wasextracted and the organic layer was dried over Na₂SO₄, filtered andconcentrated in vacuo. The residue was purified by flash columnchromatography (SiO₂, EtOAc:MeOH 10:1) to afford 80 (13 mg, 74%) as awhite solid.

Rf: 0.37 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 7.23–7.11 (m, 3H), 7.06–7.01 (m, 2H), 6.43 (s,1H), 5.95 (d, J=1.2 Hz, 1H), 5.88 (d, J=1.2 Hz, 1H), 5.71 (bs, 1H), 5.19(bs, 1H), 4.45 (d, J=3.0 Hz, 1H), 4.37 (bs, 1H), 4.02–3.96 (m, 1H),3.75–3.68 (m, 2H), 3.48 (s, 3H), 3.41–3.36 (m, 2H), 3.28–3.24 (m, 1H),3.15 (d, J=7.5 Hz, 1H), 3.01–2.88 (m, 2H), 2.70 (d, J=15.9 Hz, 1H),2.57–2.51 (m, 2H), 2.31 (s, 3H), 2.27 (s, 3H), 2.00 (s, 6H), 1.77–1.68(m, 1H).

ESI-MS m/z: Calcd. for C₃₆H₄₁N₃O₈: 643.3. Found (M−17)⁺: 626.2.

Example 75

To a solution of 73 (23 mg, 0.036 mmol) in CH₃CN/H₂O (1.5 mL/1 mL),AgNO₃ (183 mg, 1.08 mmol) was added and the reaction was stirred at 23°C. for 24 h. Then, brine (10 mL) and Aq sat NaHCO₃ (10 mL) were added at0° C. and the mixture was stirred for 15 min, filtered through a pad ofcelite and washed with CH₂Cl₂ (20 mL). The solution was extracted andthe organic layer was dried over Na₂SO₄, filtered and concentrated invacuo. The residue was purified by flash column chromatography (SiO₂,gradient EtOAc:MeOH 5:1 to MeOH) to afford 81 (9.3 mg, 41%) as a whitesolid.

Rf: 0.3 (EtOAc:MeOH 5:1).

¹HNMR (300 MHz, CDCl₃) δ 7.17–7.13 (m, 3H), 6.85 (m, 2H), 6.54 (s, 1H),5.90 (d, J=1.5 Hz, 1H), 5.84 (d, J=1.5 Hz, 1H), 5.22 (m, 1H), 4.43 (bs,1H), 4.39 (d, J=2.4 Hz, 1H), 4.00 (d, J=2.4 Hz, 1H), 3.71 (s, 3H),3.64–3.29 (m, 2H), 3.16 (d, J=8.7 Hz, 1H), 2.98–2.88 (m, 3H), 2.67 (d,J=14.8 Hz, 1H), 2.45 (d, J=18.3 Hz, 1H), 2.33 (s, 3H), 2.28 (s, 3H),2.22 (s, 3H), 1.97 (s, 3H), 1.68 (dd, J₁=12.8 Hz, J₂=14.7 Hz, 1H).

ESI-MS m/z: Calcd. for C₃₅H₃₉N₃O₈: 629.7. Found (M⁺—OH): 612.3.

Example 76

To a solution of 74 (20 mg, 0.0346 mmol) in CH₃CN/H₂O (1.5 mL/1 mL),AgNO₃ (176.6 mg, 1.04 mmol) was added and the reaction was stirred at23° C. for 24 h. Then, brine (10 mL) and Aq sat NaHCO₃ (10 mL) wereadded at 0° C. and the mixture was stirred for 15 min, filtered througha pad of celite and washed with CH₂Cl₂ (20 mL). The solution wasextracted and the organic layer was dried over Na₂SO₄, filtered andconcentrated in vacuo. The residue was purified by flash columnchromatography (SiO₂, EtOAc:MeOH 1:1) to afford 82 (12.9 mg, 66%) as awhite solid.

Rf: 0.3 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 6.50 (s, 1H), 5.95 (d, J=1.2 Hz, 1H), 5.89 (d,J=1.2 Hz, 1H), 5.19 (d, 1H), 4.46 (d, J=3.0 Hz, 1H), 4.38 (d, J=1.8 Hz,1H), 4.00 (d, J=2.1 Hz, 1H), 3.74 (s, 3H), 3.70–3.66 (m, 1H), 3.38 (dt,J₁=2.7 Hz, J₂=13.2 Hz, 1H), 3.25 (d, J=13.8 Hz, 1H), 3.16 (d, J=7.5 Hz,1H), 2.96 (dd, J₁=7.2 Hz, J₂=17.7 Hz, 1H), 2.71 (d, J=15.6 Hz, 1H), 2.40(d, J=18.0 Hz, 1H), 2.30 (s, 3H), 2.29 (s, 3H), 2.24 (s, 3H), 1.97 (s,3H), 1.71 (dd, J₁=11.7 Hz, J₂=15.3 Hz, 1H), 1.60–1.48 (m, 2H), 0.67 (t,J=7.5 Hz, 3H).

ESI-MS m/z: Calcd. for C₃₀H₃₇N₃O₈: 567.6. Found (M−17)⁺: 550.2.

Example 77

To a solution of 77 (14 mg, 0.0226 mmol) in CH₃CN/H₂O (1.5 mL/1 mL).AgNO₃ (115.3 mg, 0.68 mmol) was added and the reaction was stirred at23° C. for 24 h. Then, brine (10 mL) and Aq sat NaHCO₃ (10 mL) wereadded at 0° C. and the mixture was stirred for 15 min, filtered througha pad of celite and washed with CH₂Cl₂ (15 mL). The solution wasextracted and the organic layer was dried over Na₂SO₄, filtered andconcentrated in vacuo. The residue was purified by flash columnchromatography (SiO₂, EtOAc:MeOH 5:1) to afford 83 (9 mg, 65%) as awhite solid.

Rf: 0.25 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 6.50 (s, 1H), 5.96 (d, J=1.5 Hz, 1H), 5.89 (d,J=1.5 Hz, 1H), 5.73 (bs, 1H), 4.44 (d, J=3.6 Hz, 1H), 4.37 (s, 1H), 4.01(d, J=2.4 Hz, 1H), 3.77 (s, 3H), 3.73–3.64 (m, 1H), 3.39 (dt, J₁=3.0 Hz,J₂=9.3 Hz, 1H), 3.22 (d, J=14.5 Hz, 1H), 3.16 (d, J=7.5 Hz, 1H), 2.95(dd, J₁=8.1 Hz, J₂=17.4 Hz, 1H), 2.70 (d, J=14.5 Hz, 1H), 2.41 (d,J=18.3 Hz, 1H), 2.30 (s, 3H), 2.29 (s, 3H), 2.25 (s, 3H), 1.96 (s, 3H),1.71 (dd, J₁=12.0 Hz, J₂=15.6 Hz, 1H), 1.48–1.46 (m, 2H), 1.24–1.10 (m,4H), 1.00–0.95 (m, 2H), 0.80 (t, J=7.2 Hz, 3H).

ESI-MS m/z: Calcd. for C₃₃H₄₃N₃O₈: 609.7. Found (M−17)⁺: 592.3.

Example 78

To a solution of 78 (15 mg, 0.025 mmol) in CH₃CN/H₂O (1.5 mL/1 mL),AgNO₃ (130 mg, 0.764 mmol) was added and the reaction was stirred at 23°C. for 24 h. Then, brine (10 mL) and Aq sat NaHCO₃ (10 mL) were added at0° C. and the mixture was stirred for 15 min, filtered through a pad ofcelite and washed with CH₂Cl₂ (15 mL). The solution was extracted andthe organic layer was dried over Na₂SO₄, filtered and concentrated invacuo. The residue was purified by flash column chromatography (SiO₂,gradient EtOAc to EtOAc:MeOH 1:1) to afford 84 (10 mg, 71%) as a whitesolid.

Rf: 0.19 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 6.49 (s, 1H), 6.47–6.37 (m, 1H), 5.94 (d,J=1.5 Hz, 1H), 5.88 (d, J=1.5 Hz, 1H), 5.77 (bs, 1H), 5.26 (d, J=5.7 Hz,1H), 4.93 (d, J=14.7 Hz, 1H), 4.48 (d, J=11.1 Hz, 1H), 4.38 (d, J=2.7Hz, 1H), 4.02 ((d, J=2.1 Hz, 1H), 3.79 (s, 3H), 3.76–3.72 (m, 1H), 3.42(dt, J₁=2.7 Hz, J₂=12.0 Hz, 1H), 3.28 (d, J=13.2 Hz, 1H), 3.15 (d, J=6.6Hz, 1H), 2.96 (dd, J₁=8.7 Hz, J₂=18.0 Hz, 1H), 2.70 (d, J=15.0 Hz, 1H),2.38 (d, J=18.0 Hz, 1H), 2.30 (s, 3H), 2.28 (s, 3H), 1.95 (s, 3H), 1.72(dd, J₁=12.3 Hz, J₂=17.4 Hz, 1H), 1.98 (dd, J₁=1.5 Hz, J₂=6.9 Hz, 3H).

ESI-MS m/z: Calcd. for C₃₁H₃₇N₃O₈: 579.6. Found (M−17)⁺: 562.3.

Example 79

To a solution of 43 (25 mg, 0.422 mmol) in CH₂Cl₂ (0.3 mL),hydrocinnamoyl chloride (6.27 μL, 0.422 mmol) and pyridine (3.41 mL,0.422 mmol) were added at 0° C. The reaction mixture was stirred for 1 hand then, the solution was diluted with CH₂Cl₂ (10 mL) and washed with0.1 N HCl (5 mL). The organic layer was dried over Na₂SO₄, filtered, andthe solvent was eliminated under reduced pressure. The residue waspurified by flash column chromatography (SiO₂, gradient Hex: EtOAc 4:1to EtOAc) to afford 85 (30 mg, 68%) as a white solid.

Rf: 0.54 (EtOAcMeOH 10:1).

¹H NMR (300 MHz, CDCl₃) δ 7.28–7.14 (m, 5H), 6.45 (s, 1H), 6.07 (brd,1H), 5.99 (d, J=1.2 Hz, 1H), 5.90 (d, J=1.2 Hz, 1H), 5.88 (s, 1H), 5.31(brt, 1H), 4.09–4.06 (m, 3H), 3.80–3.75 (m, 1H), 3.73 (s, 3H), 3.57–3.51(m, 2H), 3.38 (d, J=7.5 Hz, 1H), 3.24 (m, 1H), 3.00 (dd, J₁=8.4 Hz,J₂=18.0 Hz, 1H), 2.89–2.85 (m, 2H), 2.79 (d, J=16.5 Hz, 1H), 2.61 (d,J=18.0 Hz, 1H), 2.31 (s, 3H), 2.28 (s, 3H), 2.22 (s, 3H), 2.00 (s, 3H),1.79 (dd, J₁=12.3 Hz, J₂=16.2 Hz, 1H), 0.72 (d, J=6.6 Hz, 3H).

ESI-MS m/z: Calcd. for C₄₀H₄₅N₅O₈: 723.8. Found (M+23)⁺: 746.3.

Example 80

To a solution of 43 (20 mg, 0.0338 mmol) in CH₂Cl₂ (0.25 mL), hexanoylchloride (4.72 μL, 0.0338 mmol) and pyridine (2.73 μL, 0.0338 mmol) wereadded at 0° C. The reaction mixture was stirred for 1 h and then, thesolution was diluted with CH₂Cl₂ (10 mL) and washed with 0.1 N HCl (5mL). The organic layer was dried over Na₂SO₄, filtered, and the solventwas eliminated under reduced pressure. The residue was purified by flashcolumn chromatography (SiO₂, gradient Hex:EtOAc 1:1 to EtOAc) to afford86 (10 mg, 43%) as a white solid.

Rf: 0.74 (EtOAc:MeOH 10:1).

¹H NMR (300 MHz, CDCl₃) δ 6.47 (s, 1H), 6.12 (brd, 1H), 6.00 (d, J=1.2Hz, 1H), 5.91 (d, J=1.2 Hz, 1H), 5.30 (m, 1H), 4.09–3.99 (m, 3H),3.84–3.82 (m, 1H), 3.75 (s, 3H), 3.57–3.55 (m, 2H), 3.39 (d, J=6.9 Hz,1H), 3.24 (d, J=12.0 Hz, 1H), 3.04 (dd, J₁=9.0 Hz, J₂=18.3 Hz, 1H), 2.77(d, J=15.3 Hz, 1H), 2.63 (d, J=18.0 Hz, 1H), 2.32 (s, 3H), 2.28 (s, 3H),2.25 (s, 3H), 2.00 (s, 3H), 1.80 (dd, J₁=11.7 Hz, J₂=15.6 Hz, 1H),1.55–1.50 (m, 2H), 1.30–1.22 (m, 6H), 0.87 (t, J=6.9 Hz, 3H), 0.75 (d,J=6.6 Hz, 3H).

ESI-MS m/z: Calcd. for C₃₇H₄₇N₅O₈: 689.8. Found (M+1)⁺: 690.3.

Example 81

To a solution of 43 (33 mg, 0.0557 mmol) in CH₂Cl₂ (0.4 mL), phenylacetyl chloride (7.36 μL, 0.0557 mmol) and pyridine (4.5 μL, 0.0557mmol) were added at 0° C. The reaction mixture was stirred for 1 h andthen, the solution was diluted with CH₂Cl₂ (10 mL) and washed with 0.1 NHCl (5 mL). The organic layer was dried over Na₂SO₄, filtered, and thesolvent was eliminated under reduced pressure. The residue was purifiedby flash column chromatography (SiO₂, gradient Hex:EtOAc 2:1) to afford87 (13 mg, 32%) as a white solid.

Rf: 0.63 (Hex:EtOAc:MeOH 5:10:2).

¹H NMR (300 MHz, CDCl₃) δ 7.37–7.20 (m, 5H), 6.26 (s, 1H), 6.14 (d,J=6.6 Hz, 1H), 5.98 (d, J=1.2 Hz, 1H), 5.83 (s, 1H), 5.27 (t, J=6.2 Hz,1H), 4.11 (d, J=2.1 Hz, 1H), 4.07 (d, J=3.0 Hz, 1H), 4.04 (s, 1H),3.86–3.81 (m, 1H), 3.70 (s, 3H), 3.54–3.53 (m, 2H), 3.44 (bs, 2H), 3.36(d, J=8.1 Hz, 1H), 3.22 (dt, J₁=2.7 Hz, J₂=12.0 Hz, 1H), 2.93 (dd,J₁=7.2 Hz, J₂=18.3 Hz, 1H), 2.77 (d, J=14.4 Hz, 1H), 2.59 (d, J=18.0 Hz,1H), 2.31 (s, 3H), 2.26 (s, 3H), 2.17 (s, 3H), 2.01 (s, 3H), 1.78 (dd,J₁=10.8 Hz, J₂=15.6 Hz, 1H), 0.65 (d, J=6.3 Hz, 1H).

ESI-MS m/z: Calcd. for C₃₉H₄₃N₅O₈: 709.8. Found (M+1)⁺: 710.3.

Example 82

To a solution of 43 (30 mg, 0.05 mmol) in CH₂Cl₂ (0.3 mL), propionylchloride (4.40 μL, 0.05 mmol) and pyridine (4.04 μL, 0.05 mmol) wereadded at 0° C. The reaction mixture was stirred for 1 h and then, thesolution was diluted with CH₂Cl₂ (15 mL) and washed with 0.1 N HCl (10mL). The organic layer was dried over Na₂SO₄, filtered, and the solventwas eliminated under reduced pressure. The residue was purified by flashcolumn chromatography (SiO₂, gradient Hex:EtOAc 1:1 to EtOAc) to afford88 (18 mg, 56%) as a white solid.

Rf: 0.49 (Hex:EtOAc:MeOH 1:10:2).

¹H NMR (300 MHz, CDCl₃) δ 6.46 (s, 1H), 6.16 (brd, 1H), 5.99 (d, J=1.2Hz, 1H), 5.95 (s, 1H), 5.90 (d, J=1.2 Hz, 1H), 5.34 brt, 1H), 4.12–4.06(m, 3H), 3.84 (bs, 1H), 3.74 (s, 3H), 3.63 (dd, J₁=6.3 Hz, J₂=212.9 Hz,1H), 3.50–3.48 (m, 1H), 3.39 (d, J=8.1 Hz, 1H), 3.23 (d, J=11.7 Hz, 1H),3.00 (dd, J₁=8.4 Hz, J₂=18.3 Hz, 1H), 2.78 (d, J=15.6 Hz, 1H), 2.63 (d,J=18.3 Hz, 1H), 2.31 (s, 3H), 2.27 (s, 3H), 1.87–1.80 (m, 1H), 1.06 (t,J=7.5 Hz, 3H), 0.74 (d, J=6.9 Hz, 3H).

ESI-MS m/z: Calcd. for C₃₄H₄₁N₅O₈: 647.7. Found (M+1)⁺: 648.2.

Example 83

To a solution of 43 (20 mg, 0.0338 mmol) in CH₂Cl₂ (0.3 mL), propionylchloride (3.238 μL, 0.0338 mmol) and pyridine (2.73 μL, 0.0338 mmol)were added at 0° C. The reaction mixture was stirred for 1 h and then,the solution was diluted with CH₂Cl₂ (10 mL) and washed with 0.1 N HCl(5 mL). The organic layer was dried over Na₂SO₄ filtered, and thesolvent was eliminated under reduced pressure. The residue was purifiedby flash column chromatography (SiO₂, gradient Hex:EtOAc 3:1 to AcOEt)to afford 89 (11.5 mg, 52%) as a white solid.

Rf: 0.57 (EtOAc:MeOH 10:1).

¹H NMR (300 MHz, CDCl₃) δ 6.82–6.70 (m, 1H), 6.46 (s, 1H), 6.11 (d, 1H),6.00 (d, J=1.5 Hz, 1H), 5.89 (d, J=1.5 Hz, 1H), 5.85 (s, 1H), 5.77 (dd,J₁=1.5 Hz, J₂=15.3 Hz, 1H), 5.37 (bst, 1H), 4.13–4.06 (m, 3H), 3.19 (m,1H), 3.73 (s, 3H), 3.55 (m, 2H), 3.38 (d, J=1.5 Hz, 1H), 3.23 (d, J=11.4Hz, 1H), 3.00 (dd, J₁=8.4 Hz, J₂=18.3 Hz, 1H), 2.78 (d, J=15.0 Hz, 1H),2.65 (d, J=18.0 Hz, 1H), 2.31 (s, 3H), 2.28 (s, 3H), 2.22 (s, 3H), 2.00(s, 3H), 1.85–1.82 (m, 4H), 0.77 (d, J=6.3 Hz, 3H).

ESI-MS m/z: Calcd. for C₃₅H₄₁N₅O₈: 659.7. Found (M+1)⁺: 660.3.

Example 84

To a solution of 43 (15 mg, 0.0253 mmol) in CH₂Cl₂ (0.3 mL),Cbz-L-Val-OH (6.39 mg, 0.0253 mmol) and carbonyl dimidazole (4.86 mg,0.03 mmol) were added at 0° C. The reaction mixture was stirred for 16 hat room temperature and then, the solution was diluted with CH₂Cl₂ (15mL) and washed with 0.1 N HCl (10 mL). The organic layer was dried overNa₂SO₄, filtered, and the solvent was eliminated under reduced pressure.The residue was purified by flash column chromatography (SiO₂, gradientHex:EtOAc 1:1 to EtOAc) to afford 90 (6.7 mg, 32%) as a white solid.

Rf: 0.79 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 7.35 (bs, 5H), 6.46 (s, 1H), 6.28 (d, J=6.0Hz, 1H), 5.98 (d, J=1.2 Hz, 1H), 5.89 (d, J=1.2 Hz, 1H), 5.77 (s, 1H),5.44 (bs, 1H), 5.30 (bs, 1H), 5.08 (s, 2H), 4.09–4.06 (m, 3H), 3.94–3.89(m, 1H), 3.70–3.66 (m, 5H), 3.38 (d, J=11.7 Hz, 1H), 3.01 96 (dd, J₁=7.8Hz, J₂₌18.3 Hz, 1H), 2.79 (d, J=14.1 Hz, 1H), 2.63 (d, J=18.0 Hz, 1H),2.30 (s, 3H), 2.28 (s, 3H), 2.20 (s, 3H), 1.99 (s, 3H9, 1.97–1.81 (m,2H), 0.83 (d, J=6.6 Hz, 3H), 0.80 (d, J=6.6 Hz, 3H), 0.75 (d, J=6.9 Hz,3H).

ESI-MS m/z: Calcd. for C₄₄H₅₂N₆O₁₀: 824.9. Found (M+1)⁺: 825.4.

Example 85

To a solution of 62 (20 mg, 0.030 mmol) in CH₃CN/H₂O (1.5 mL/1 mL),AgNO₃ (154 mg, 0.90 mmol) was added and the reaction was stirred at 23°C. for 24 h. Then, brine (10 mL) and Aq sat NaHCO₃ (10 mL) were added at0° C. and the mixture was stirred for 15 min, filtered through a pad ofcelite and washed with CH₂Cl₂ (15 mL). The solution was extracted andthe organic layer was dried over Na₂SO₄, filtered and concentrated invacuo. The residue was purified by flash column chromatography (SiO₂,gradient EtOAc to EtOAc:MeOH 3:1) to afford 91 (13 mg, 66%) as a whitesolid.

Rf: 0.18 (EtOAc:MeOH 10:1).

¹H NMR (300 MHz, CDCl₃) δ 6.49 (s, 1H), 6.16 (d, 1H), 5.98 (d, J=1.5 Hz,1H), 5.89 (d, J=1.5 Hz, 1H), 5.32 (bs, 1H), 4.41 (bs, 1H), 4.00 (bs,1H), 3.79 (s, 3H), 3.70–3.65 (m, 2H), 3.37–3.32 (m, 2H), 3.19–3.17 (m,1H), 2.94 (dd, J₁=9.0 Hz, J₂=15.0 Hz, 1H), 2.74 (d, J=15.9 Hz, 1H), 2.46(d, J=17.1 Hz, 1H), 2.31 (s, 3H), 2.28 (s, 3H), 2.27 (s, 3H), 2.04–2.01(m, 2H), 1.98 (s, 3H), 1.64–1.62 (m, 1H), 1.54–1.52 (m, 2H), 0.89–0.84(m, 6H).

ESI-MS m/z: Calcd. for C₃₄H₄₄N₄O₉: 652.7. Found (M−17)⁺: 635.3.

Example 86

To a solution of 85 (10 mg, 0.0138 mmol) in CH₃CN/H₂O (1.5 mL/1 mL),AgNO₃ (70.4 mg, 0.414 mmol) was added and the reaction was stirred at23° C. for 24 h. Then, brine (10 mL) and Aq sat NaHCO₃ (10 mL) wereadded at 0° C. and the mixture was stirred for 15 min, filtered througha pad of celite and washed with CH₂Cl₂ (15 mL). The solution wasextracted and the organic layer was dried over Na₂SO₄, filtered andconcentrated in vacuo. The residue was purified by flash columnchromatography (SiO₂, gradient EtOAc to EtOAc:MeOH 4:1) to afford 92 (7mg, 71%) as a white solid.

Rf: 0.20 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 7.25–7.13 (m, 5H), 6.47 (s, 1H), 6.13 (brd,1H) 5.97 (d, J=1.2 Hz, 1H), 5.88 (d, J=1.2 Hz, 1H), 5.34 (brt, 1H), 4.50(bs, 1H), 4.40 (bs, 1H), 4.00 (bs, 1H), 3.76 (s, 3H), 3.70–3.65 (m, 3H),3.34 (d, J=11.7 Hz, 1H), 3.17 (d, J=5.1 Hz, 1H), 2.98–2.83 (m, 3H), 2.72(d, J=14.4 Hz, 1H), 2.44 (d, J=19.2 Hz, 1H), 2.30 (s, 3H), 2.27 (s, 6H),1.97 (s, 3H), 1.72 (m, 1H), 0.82 (d, J=6.6 Hz, 3H).

ESI-MS m/z: Calcd. for C₃₉H₄₆N₄O₉: 714.8. Found (M−17)⁺: 697.3.

Example 87

To a solution of 86 (6 mg, 0.0087 mmol) in CH₃CN/H₂O (1.5 mL/1 mL),AgNO₃ (44 mg, 0.26 mmol) was added and the reaction was stirred at 23°C. for 24 h. Then, brine (10 mL) and Aq sat NaHCO₃ (10 mL) were added at0° C. and the mixture was stirred for 15 min. filtered through a pad ofcelite and washed with CH₂Cl₂ (15 mL). The solution was extracted andthe organic layer was dried over Na₂SO₄, filtered and concentrated invacuo. The residue was purified by flash column chromatography (SiO₂,gradient EtOAc to EtOAc:MeOH 5:1) to afford 93 (5 mg, 85%) as a whitesolid.

Rf: 0.018 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 6.48 (s, 1H), 6.17 (d, 1H), 5.98 (d, J=1.5 Hz,1H), 5.89 (d, J=1.5 Hz, 1H), 5.33 (bs, 1H), 4.51 (d, 1H), 4.40 (d, 1H),4.00 (d, 1H), 3.78 (s, 3H), 3.76–3.65 (m, 2H), 3.36–3.32 (m, 2H), 3.18(d, J=6.9 Hz, 1H), 2.98–2.89 (m, 1H), 2.71 (d, J=15.0 Hz, 1H), 2.45 (d,J=17.7 Hz, 1H), 2.31 (s, 3H), 2.27 (s, 3H), 2.26 (s, 3H), 1.98 (s, 3H),1.68–1.50 (m, 3H), 1.29–1.19 (m, 6H), 0.88–0.84 (m, 6H).

ESI-MS m/z: Calcd. for C₃₆H₄₈N₄O₉: 680.7. Found (M−17)⁺: 663.3.

Example 88

To a solution of 87 (12 mg, 0.0169 mmol) in CH₃CN/H₂O(1.5 mL/1 mL),AgNO₃ (86 mg, 0.507 mmol) was added and the reaction was stirred at 23°C. for 24 h. Then, brine (10 mL) and Aq sat NaHCO₃ (10 mL) were added at0° C. and the mixture was stirred for 15 min, filtered through a pad ofcelite and washed with CH₂Cl₂ (15 mL). The solution was extracted andthe organic layer was dried over Na₂SO₄, filtered and concentrated invacuo. The residue was purified by flash column chromatography (SiO₂,gradient EtOAc to EtOAc:MeOH 5:1) to afford 94 (8.8 mg, 74%) as a whitesolid.

Rf: 0.28 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 7.34–7.18 (m, 5H), 6.37 (s, 1H), 6.20 (d, 1H),5.96 (d, J=1.5 Hz, 1H), 5.88 (d, J=1.5 Hz, 1H), 5.30 (t, 1H), 4.50 (bs,1H), 4.39 (d, J=1.8 Hz, 1H), 3.99 (d, J=2.1 Hz, 1H), 3.73 (s, 3H),3.69–3.60 (m, 3H), 3.37–3.30 (m, 3H), 3.17 (d, J=18.1 Hz, 1H), 2.89 (dd,J₁=7.5 Hz, J₂=18.3 Hz, 1H), 2.31 (s, 3H), 2.25 (s, 3H), 2.21 (s, 3H),1.99 (s, 3H), 1.71 (dd, J₁=11.7 Hz, J₂=15.0 Hz, 1H), 0.77 (d, J=6.6 Hz,1H).

ESI-MS m/z: Calcd. for C₃₈H₄N₄O₉: 700.7. Found (M−17)⁺: 683.2.

Example 89

To a solution of 88 (14 mg, 0.0216 mmol) in CH₃CN/H₂O (1.5 mL/1 mL),AgNO₃ (110 mg, 0.648 mmol) was added and the reaction was stirred at 23°C. for 24 h. Then, brine (10 mL) and Aq sat NaHCO₃ (10 mL) were added at0° C. and the mixture was stirred for 15 min, filtered through a pad ofcelite and washed with CH₂Cl₂ (15 mL). The solution was extracted andthe organic layer was dried over Na₂SO₄, filtered and concentrated invacuo. The residue was purified by flash column chromatography (SiO₂,gradient EtOAc to EtOAc:MeOH 5:1) to afford 95 (9.7 mg, 70%) as a whitesolid.

Rf: 0.16 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 6.48 (s, 1H), 6.10 (d, 1H), 5.97 (d, J=1.2 Hz,1H), 5.89 (d, J=1.2 Hz, 1H), 5.36 (bs, 1H), 4.51 (bs, 1H), 4.40 (d,J=2.1 Hz, 1H), 4.00 (d, J=2.1 Hz, 1H), 3.78 (s, 3H), 3.76–3.62 (m, 3H),3.33 (d, J=11.7 Hz, 1H), 3.18 (d, J=8.4 Hz, 1H), 2.94 (dd, J₁=8.4 Hz,J₂=16.5 Hz, 1H), 2.72 (d, J=15.0 Hz, 1H), 2.45 (d, J=18.3 Hz, 1H), 2.31(s, 3H), 2.27 (s, 3H), 2.22 (s, 3H), 1.97 (s, 3H), 1.86 (m, 2H), 1.73(dd, J₁=12.0 Hz, J₂₌15.0 Hz, 1H), 1.05 (t, J=7.8 Hz, 3H), 0.83 (d, J=6.9Hz, 3H).

ESI-MS m/z: Calcd. for C₃₃H₄₂N₄O₉: 638.7. Found (M−17)⁺: 621.2.

Example 90

To a solution of 89 (10 mg, 0.015 mmol) in CH₃CN/H₂O (155 mL/1 mL).AgNO₃ (77.2 mg, 0.454 mmol) was added and the reaction was stirred at23° C. for 24 h. Then, brine (10 mL) and Aq sat NaHCO₃ (10 mL) wereadded at 0° C. and the mixture was stirred for 15 min, filtered througha pad of celite and washed with CH₂Cl₂ (15 mL). The solution wasextracted and the organic layer was dried over Na₂SO₄, filtered andconcentrated in vacuo. The residue was purified by flash columnchromatography (SiO₂, gradient EtOAc to EtOAc:MeOH 1:1) to afford 96 (9mg, 92%) as a white solid.

Rf: 0.016 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 6.76–6.69 (m, 1H), 6.47 (s, 1H), 6.18 (brd,1H), 5.97 (d, J=1.5 Hz, 1H), 5.88 (d, J=1.5 Hz, 1H), 5.71 (dd, J₁=1.5Hz, J₂=16.2 Hz, 3H), 5.32 (bs, 1H), 4.50 (m, 1H), 4.41 (m, 1H), 3.99 (m,1H), 3.78 (m, 4H), 3.64–3.58 (m, 2H), 3.34 (d, J=11.1 Hz, 1H), 3.17 (d,J=8.6 Hz, 1H), 2.95 (dd, J₁=7.5 Hz, J₂=17.4 Hz, 1H), 2.70 (d, J=16.2 Hz,1H), 2.48 (d, J=17.7 Hz, 1H), 2.31 (s, 3H), 2.27 (s, 3H), 2.17 (s, 6H),1.97 (s, 3H), 1.82–1.74 (m, 4H), 0.88 (t, J=5.2 Hz, 3H).

ESI-MS m/z: Calcd. for C₃₄H₄₂N₄O₉: 650.7. Found (M−17)⁺: 633.3.

Example 91

To a solution of 25 (100 mg, 0.177 mmol) in CH₂Cl₂ (0.5 mL), butyrylchloride (24 μL, 0.23 mmol) and pyridine (17 μL, 0.212 mmol) were addedat 0° C. The reaction mixture was stirred for 2 h at room temperatureand then, the solution was diluted with CH₂Cl₂ (30 mL) and washed with0.1 N HCl (20 mL). The organic layer was dried over Na₂SO₄, filtered,and the solvent was eliminated under reduced pressure. The residue waspurified by flash column chromatography (SiO₂, Hex:EtOAc 3:1) to afford97 (99 mg, 88%) as a colorless oil.

Rf: 0.64 (Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 6.66 (s, 1H), 6.16–6.05 (m, 1H), 5.93 (d,J=1.2 Hz, 1H), 5.87 (d, J=1.2 Hz, 1H), 5.40 (dd, J₁=1.2 Hz, J₂=17.1 Hz1H), 5.26 (dd, J₁=1.2 Hz, J₂=10.2 Hz, 1H), 5.13–5.08 (m, 2H), 4.44 (dd,J₁=3.6 Hz, J₂=11.1 Hz, 1H), 4.21–4.07 (m, 5H), 3.74 (m, 1H), 3.72 (s,1H), 3.57 (s, 3H), 3.35 (d, J=10.5 Hz, 1H), 3.26–3.21 (m, 2H), 3.98 (dd,J₁=8.7 Hz, J₂=18.0 Hz, 1H), 2.54 (d, J=18.0 Hz), 2.30 (s, 3H), 2.21 (s,3H), 2.13 (s, 3H), 1.92–1.65 (m, 3H), 1.42–1.34 (m, 2H), 0.80 (t, J=7.5Hz, 3H).

ESI-MS m/z: Calcd. for C₃₅H₄₃N₃O₉: 633.7. Found (M+1)⁺: 634.3.

Example 92

To a solution of 25 (100 mg, 0.177 mmol) in CH₂Cl₂ (0.4 mL),trans-3-(trifluoromethyl)cinnamoyl chloride (35 μL, 0.23 mmol) andpyridine (17 μL, 0.212 mmol) were added at 0° C. The reaction mixturewas stirred for 1 h at room temperature and then, the solution wasdiluted with CH₂Cl₂ (30 mL) and washed with 0.1 N HCl (20 mL). Theorganic layer was dried over Na₂SO₄, filtered, and the solvent waseliminated under reduced pressure. The residue was purified by flashcolumn chromatography (SiO₂, gradient Hex:EtOAc 6:1 to Hex:EtOAc 1:1) toafford 98 (122 mg, 90%) as a white solid. Rf: 0.478 (Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 7.64–7.48 (m, 4H), 7.37 (d, J=15.6 Hz, 1H),6.62 (s, 1H), 6.16–6.07 (m, 1H), 6.12 (d, J=15.6 Hz, 1H), 5.94 (d, J=1.2Hz, 1H), 5.89 (d, J=1.2 Hz, 1H), 5.41 (dd, J₁=1.8 Hz, J₂=17.1 Hz, 1H),5.28 (dd, J₁=1.8 Hz, J₂=12.0 Hz, 1H), 5.04 (q, J=6.0 Hz, 1H), 4.60 (dd,J₁=3.3 Hz, J₂=11.1 Hz, 1H), 4.22–4.15 (m, 5H), 3.90 (dd, J₁=4.2 Hz,J₂=11.1 Hz, 1H), 3.55 (s, 3H), 3.38 (s, 3H), 3.35–3.34 (m, 1H),3.27–3.25 (m, 1H), 3.22 (bs, 1H), 2.98 (dd, J₁=7.8 Hz, J₂=18.0 Hz, 1H),2.61 (d, J=17.7 Hz, 1H), 2.29 (s, 3H), 2.16 (s, 3H), 2.00 (s, 3H), 1.80(dd, J₁=11.7 Hz, J₂=15.6 Hz, 1H).

ESI-MS m/z: Calcd. for C₄₁H₂F₃N₃O: 761.7. Found (M+1)⁺: 762.3.

Example 93

To a solution of 25 (68 mg, 0.12 mmol) in CH₂Cl₂ (0.4 mL),hydrocynnamoyl chloride (20 μL, 1.12 mmol) and pyridine (10 μL, 1.01mmol) were added at 0° C. The reaction mixture was stirred for 2 h atroom temperature and then, the solution was diluted with CH₂Cl₂ (30 mL)and washed with 0.1 N HCl (20 mL). The organic layer was dried overNa₂SO₄, filtered, and the solvent was eliminated under reduced pressure.The residue was purified by flash column chromatography (SiO₂, gradientHex:EtOAc 5:1 to Hex:EtOAc 2:1) to afford 99 (41 mg, 49%) as a whitesolid. Rf: 0.47 (Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 7.29–7.18 (m, 3H), 7.04–7.02 (m, 2H), 6.66 (s,1H), 6.16–6.07 (m, 1H), 5.93 (d, J=1.2 Hz, 1H), 5.87 (d, J=1.2 Hz, 1H),5.40 (dd, J₁=1.7 Hz, J₂=17.4 Hz, 1H), 5.26 (dd, J₁=1.7 Hz, J₂=10.2 Hz,1H), 5.09 (dd, J₁=6.0 Hz, J₂=8.7 Hz, 2H), 4.43 (dd, J₁=3.3 Hz, J₂=11.1Hz, 1H), 4.20–4.14 (m, 3H), 4.06 (t, J=3.7 Hz, 1H), 4.02 (d, J=2.4 Hz,1H), 3.72 (dd, J₁=4.5 Hz, J₂=11.1 Hz, 1H), 3.56 (s, 3H), 3.55 (s, 3H),3.32 (brd, J=8.7 Hz, 1H), 3.26 (dd, J₁=1.9 Hz, J₂=8.1 Hz, 1H), 3.23–3.20(m, 1H), 3.01 (brd, J=8.1 Hz, 1H), 3.23–3.20 (m, 1H), 3.26 (dd, J₁=1.9Hz, J₂=8.1 Hz, 1H), 2.95 (d, J=1.8 Hz, 1H), 2.71–2.64 (m, 3H), 2.53 (d,J=17.7 Hz, 1H), 2.26 (s, 3H), 2.14 (s, 6H), 1.83 (dd, J₁=12.3 Hz,J₂=15.9 Hz, 1H).

ESI-MS m/z: Calcd. for C₄₀H₄₅F₃N₃O₈: 695.3. Found (M+1)⁺: 696.3.

Example 94

To a solution of 25 (100 mg, 0.177 mmol) in CH₂Cl₂ (0.4 mL), cynnamoylchloride (35 mg, 0.21 mmol) and pyridine (17 μL, 0.21 mmol) were addedat 0° C. The reaction mixture was stirred for 2 h at room temperatureand then, the solution was diluted with CH₂Cl₂ (30 mL) and washed with0.1 N HCl (20 mL). The organic layer was dried over Na₂SO₄, filtered,and the solvent was eliminated under reduced pressure. The residue waspurified by flash column chromatography (SiO₂, Hex:EtOAc 6:1) to afford100 (94 mg, 76%) as a white solid. Rf: 0.49 (Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 7.42–7.33 (m, 6H), 6.62 (s, 1H), 6.16–6.05 (m,1H), 6.10 (d, J=15.9 Hz, 1H), 5.94 (d, J=1.2 Hz, 1H), 5.88 (d, J=1.2 Hz,1H), 5.43 (dd, J₁=3.0 Hz, J₂=17.1 Hz, 1H), 5.27 (dd, J₁=3.0 Hz, J₂=12.0Hz, 1H), 5.04 (q, J=6.0 Hz, 1H) 4.55 (dd, J₁=3.9 Hz, J₂=11.1 Hz, 1H),4.22–4.15 (m, 5H), 3.87 (dd, J₁=4.5 Hz, J₂=11.1 Hz, 1H), 3.55 (s, 3H),3.39 (s, 3H), 3.36–3.33 (m, 1H), 3.26–3.22 (m, 2H), 2.98 (dd, J₁=8.1 Hz,J₂=17.7 Hz, 1H), 2.63 (d, J=17.7 Hz, 1H), 2.29 (s, 3H), 2.03 (s, 3H),1.82 (dd, J₁=11.7 Hz, J₂=15.3 Hz, 1H).

ESI-MS m/z: Calcd. for C₄₀H₄₃N₃O₈: 693.3. Found (M+1)⁺: 694.3.

Example 95

To a solution of 97 (40 mg, 0.063 mmol) in CH₂Cl₂ (0.7 mL), acetic acid(17.8 μL), Pd(PPh₃)₂Cl₂ (3.64 mg, 0.0052 mmol) and Bu₃SnH (67.9 μL,0.252 mmol) were added at 23° C. The reaction mixture was stirred for 2h at that temperature and then, the solution was poured into a pad offlash column (SiO₂, gradient Hex:EtOAc 5:1 to Hex:EtOAc 3:1) to afford101 (30 mg, 80%) as a white solid. Rf: 0.4 (Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 6.65 (s, 1H), 5.90 (d, J=1.5 Hz, 1H), 5.82 (d,J=1.5 Hz, 1H), 5.54 (s, 1H), 5.33 (d, J=6.0 Hz, 1H), 5.13 (d, J=6.0 Hz,1H), 4.54 (dd, J₁=3.6 Hz, J₂=11.4 Hz, 1H), 4.18 (d, J=2.1 Hz, 1H), 4.13(d, J=2.4 Hz, 1H), 4.07 (t, J=3.3 Hz, 1H), 3.75 (dd, J₁=3.9 Hz, J₂=11.1Hz, 1H), 3.70 (s, 3H), 3.35 (d, J=8.4 Hz, 1H), 3.24 (dd, J₁=2.7 Hz,J₂=8.7 Hz, 1H), 3.10 (dd, J₁=2.4 Hz, J₂=15.0 Hz, 1H), 3.01 (d, J=8.1 Hz,1H), 2.95 (d, J=7.8 Hz, 1H), 2.58 (d, J=18.3 Hz, 1H), 2.29 (s, 3H), 2.21(s, 3H), 2.10 (s, 3H), 1.89–1.66 (m, 3H), 1.36–1.25 (m, 2H), 0.77 (t,J=7.5 Hz, 3H).

ESI-MS m/z: Calcd. for C₃₂H₃₉N₃O₈: 593.6. Found (M+1)⁺: 594.8

Example 96

To a solution of 98 (37 mg, 0.0485 mmol) in CH₂Cl₂ (0.7 mL), acetic acid(20 μL), Pd(PPh₃)₂Cl₂ (4 mg, 0.0057 mmol) and Bu₃SnH (53 μL, 0.194 mmol)were added at 23° C. The reaction mixture was stirred for 5 h at thattemperature and then, the solution was poured into a pad of flash column(SiO₂, gradient Hex:EtOAc 6:1 to Hex:EtOAc 2:1) to afford 102 (25 mg,71%) as a white solid. Rf: 0.38 (Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 7.63–7.60 (M, 2H), 7.50–7.49 (M, 2H), 7.24 (d,J=15.9 Hz, 1H), 6.59 (s, 1H), 5.98 (d, J=15.9 Hz, 1H), 5.92 (d, J=1.5Hz, 1H), 5.84 (d, J=1.5 Hz, 1H), 5.66 (s, 1H), 5.20 (d, J=6.0 Hz, 1H),4.87 (d, J=6.0 Hz, 1H), 4.71 (dd, J₁=2.7 Hz, J₂=10.8 Hz, 1H), 4.16–4.15(m, 3H), 3.93 (dd, J₁=3.3 Hz, J₂=11.1 Hz, 1H), 3.66 (s, 3H), 3.36 (brd,J=10.2 Hz, 1H), 3.26 (brd, J=11.7 Hz, 1H), 3.10 (brd, J=15.0 Hz, 1H),2.96 (dd, J₁=7.8 Hz, J₂=17.7 Hz, 1H), 2.62 (d, J=17.7 Hz, 1H), 2.27 (s,3H), 2.14 (s, 3H), 1.97 (s, 3H), 1.79 (dd, J₁=12.0 Hz, J₂=15.8 Hz, 1H).

ESI-MS m/z: Calcd. for C₃₈H₃₈F₃N₃O₈: 721.7. Found (M+1)⁺: 722.2.

Example 97

To a solution of 99 (41 mg, 0.059 mmol) in CH₂Cl₂ (1 mL), acetic acid(25 μL), Pd(PPh₃)₂Cl₂ (5 mg, 0.0071 mmol) and Bu₃SnH (63 μL, 0.235 mmol)were added at 23° C. The reaction mixture was stirred for 4.5 h at thattemperature and then, the solution was poured into a pad of flash column(SiO₂, gradient Hex:EtOAc 6:1 to Hex:EtOAc 1:1) to afford 103 (34.2 mg,89%) as a white solid. Rf: 0.49 (Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 7.24–7.15 (m, 3H), 7.03–7.01 (m, 2H), 6.65 (s,1H), 5.89 (bs, 1H), 5.82 (bs, 1H), 5.49(s, 1H), 5.31 (d, J=6.0 Hz, 1H),5.12 (d, J=6.0 Hz, 1H), 4.53 (dd, J₁=3.3 Hz, J₂=11.1 Hz, 1H), 4.18 (d,J=2.7 Hz, 1H), 4.07 (m, 2H), 3.75 (dd, J₁=3.9 Hz, J₂=11.1 Hz, 1H), 3.69(s, 3H), 3.62 (s, 3H), 3.32 (d, J=7.8 Hz, 1H), 3.25 (d, J=10.8 Hz, 1H),3.12 (d, J=14.7 Hz, 1H), 3.00 (d, J=7.8 Hz, 1H), 2.94 (d, J=8.1 Hz, 1H),2.66–2.60 (m, 3H), 2.57 (d, J=18.0 Hz, 1H), 2.28 (s, 3H), 2.14 (s, 3H),2.10 (bs, 3H), 1.83–1.74 (m, 1H).

ESI-MS m/z: Calcd. for C₃₇H₄₁N₃O₈: 655.7. Found (M+1)⁺: 656.3.

Example 98

To a solution of 100 (40 mg, 0.0576 mmol) in CH₂Cl₂ (1 mL), acetic acid(25 μL). Pd(PPh₃)₂Cl₂ (4.8 mg, 0.007 mmol) and Bu₃SnH (62 μL, 0.23 mmol)were added at 23° C. The reaction mixture was stirred for 5 h at thattemperature and then, the solution was poured into a pad of flash column(SiO₂, gradient Hex:EtOAc 4:1 to Hex:EtOAc 1:1) to afford 104 (30 mg,82%) as a white solid. Rf: 0.41 (Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 7.36 (s, 5H), 7.30 (d, J=16.2 Hz, 1H), 6.59(s, 1H), 5.99 (d, J=16.2 Hz, 1H), 5.91 (d, J=1.2 Hz, 1H), 5.84 (d, J=1.2Hz, 1H), 5.60 (s, 1H), 5.20 (d, J=5.6 Hz, 1H), 4.94 (d, J=5.6 Hz, 1H),4.63 (dd, J₁=3.3 Hz, J₂=11.4 Hz, 1H), 4.18–4.15 (m, 3H), 3.91 (dd,J₁=3.9 Hz, J₂=11.1 Hz, 1H), 3.66 (s, 3H), 3.49 (s, 3H), 3.35 (brd,J=15.0 Hz, 1H), 3.26 (brd, J=11.4 Hz, 1H), 3.10 (brd, J=15.0 Hz, 1H),2.96 (dd, J₁=8.4 Hz, J₂=18.0 Hz, 1H), 2.63 (d, J=18.0 Hz, 1H), 2.27 (s,3H), 2.13 (s, 3H), 2.00 (s, 3H), 1.80 (dd, J₁=12.0 Hz, J₂=14.4 Hz, 1H).

ESI-MS m/z: Calcd. for C₃₇H₃₉N₃O₈: 653.7. Found (M+23)⁺: 676.2.

Example 99

To a solution of 101 (24 mg, 0.041 mmol) in CH₂Cl₂ (0.4 mL), acetylchloride (3 μL, 0.041 mmol), and pyridine (3.3 μL, 0.041 mmol) wereadded at 0° C. The reaction mixture was stirred for 2 h and then, thesolution was diluted with CH₂Cl₂ (15 mL) and washed with 0.1 N HCl (5mL). The organic layer was dried over sodium sulphate, filtered, and thesolvent was eliminated under reduced pressure. The residue was purifiedby flash column chromatography (SiO₂, gradient Hex:EtOAc 5:1 toHex:EtOAc 1:1) to afford 105 (23 mg, 88%) as a white solid. Rf: 0.40(Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 6.66 (s, 1H), 5.97 (d, J=1.2 Hz, 1H), 5.91 (d,J=1.2 Hz, 1H), 4.58 (d, J=3.0 Hz, 1H), 4.54 (d, J=3.0 Hz, 1H), 4.07 (t,J=3.3 Hz, 1H), 3.77 (dd, J₁=3.9 Hz, J₂=11.4 Hz, 1H), 3.73 (s, 3H), 3.57(s, 3H), 3.35 (d, J=10.2 Hz, 1H), 3.22 (dt, J₁=2.7 Hz, J₂=11.7 Hz, 1H),2.98 (dd, J₁=8.1 Hz, J₂=18.0 Hz, 1H), 2.80 (d, J=13.5 Hz, 1H), 2.58 (d,J=18.0 Hz, 1H), 2.33 (s, 3H), 2.30 (s, 3H), 2.21 (s, 3H), 2.02 (s, 3H),1.89–1.76 (m, 2H) 1.72–1.66 (m, 1H), 1.37–1.25 (m, 2H), 0.78 (t, J=7.5Hz, 3H).

ESI-MS m/z: Calcd. for C₃₄H₄N₃O₉: 635.7. Found (M+1)⁺: 636.8.

Example 100

To a solution of 102 (16 mg, 0.022 mmol) in CH₂Cl₂ (0.2 mL), acetylchloride (1.9 μL, 0.0266 mmol), and pyridine (2.15 μL, 0.0266 mmol) wereadded at 0° C. The reaction mixture was stirred for 1.5 h and then, thesolution was diluted with CH₂Cl₂ (10 mL) and washed with 0.1 N HCl (7mL). The organic layer was dried over sodium sulphate, filtered, and thesolvent was eliminated under reduced pressure. The residue was purifiedby flash column chromatography (SiO₂, gradient Hex:EtOAc 4:1 toEtOAc) toafford 106 (12 mg, 71%) as a white solid. Rf: 0.60 (Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 7.83 (bs, 1H), 7.65–7.58 (m, 2H), 7.49–7.44(m, 1H), 7.14 (d, J=16.2 Hz, 1H), 6.62 (s, 1H), 6.06 (d, J=16.2 Hz, 1H),6.00 (d, J=1.2 Hz, 1H), 5.95 (d, J=1.2 Hz, 1H), 5.02 (d, J=5.7 Hz, 1H),4.96(bs, 1H), 4.92 (d, J=5.7 Hz, 1H), 4.15–4.11 (m, 3H), 3.88 (dd,J₁=3.3 Hz, J₂=11.1 Hz, 1H), 3.08 (bs, 3H), 2.93 (dd, J₁=8.1 Hz, J₂=18.3Hz, 1H), 2.80 (d, J=13.2 Hz, 1H), 2.64 (d, J=18.0 Hz, 1H), 2.31 (s, 3H),2.27 (s, 3H), 2.08 (s, 3H), 1.91 (s, 3H), 1.69 (dd, J₁=11.7 Hz, J₂=15.9Hz, 1H). ).

ESI-MS m/z: Calcd. for C₄₀H₄₀F₃N₃O₉: 763.7. Found (M+1)⁺: 764.2.

Example 101

To a solution of 103 (34 mg, 0.052 mmol) in CH₂Cl₂ (0.2 mL), acetylchloride (4.4 μL, 0.062 mmol), and pyridine (5 μL, 0.062 mmol) wereadded at 0° C. The reaction mixture was stirred for 1.5 h and then, thesolution was diluted with CH₂Cl₂ (10 mL) and washed with 0.1 N HCl (7mL). The organic layer was dried over sodium sulphate, filtered, and thesolvent was eliminated under reduced pressure. The residue was purifiedby flash column chromatography (SiO₂, gradient Hex:EtOAc 4:1 toEtOAc) toafford 107 (25.5 mg, 70%) as a white solid. Rf: 0.48 (Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 7.25–7.14 (m, 3H), 7.06–7.04 (m, 2H), 6.66 (s,1H), 5.96 (d, J=1.2 Hz, 1H), 5.91 (d, J=1.2 Hz, 1H), 5.11 (d, J=5.4 Hz,1H), 4.14 (d, J=3.3 Hz, 1H), 4.07 (d, J=3.6 Hz, 1H), 4.04 (d, J=2.7 Hz,1H), 3.78 (dd, J₁=3.3 Hz, J₂=10.8 Hz, 1H), 3.55 (s, 3H), 3.51 (s, 3H),3.33 (brd, J=8.1 Hz, 1H), 3.23 (dt, J₁=2.7 Hz, J₂=11.7 Hz, 1H), 2.97(dd, J₁=8.1 Hz, J₂=18.0 Hz, 1H), 2.81 (d, J=14.1 Hz, 1H), 2.63–2.52 (m,3H), 2.33 (s, 3H), 2.29 (s, 3H), 2.26–202 (m, 2H), 2.09 (s, 3H), 2.04(s, 3H), 1.74 (dd, J₁=12.0 Hz, J₂₌15.6 Hz, 1H).

ESI-MS m/z: Calcd. for C₃₉H₄₃N₃O₉: 697.7. Found (M+1)⁺: 698.3.

Example 102

To a solution of 104 (29 mg, 0.0443 mmol) in CH₂Cl₂ (0.3 mL), acetylchloride (3.77 μL, 0.053 mmol), and pyridine (4.3 μL, 0.053 mmol) wereadded at 0° C. The reaction mixture was stirred for 2 h and then, thesolution was diluted with CH₂Cl₂ (15 mL) and washed with 0.1 N HCl (10mL). The organic layer was dried over sodium sulphate, filtered, and thesolvent was eliminated under reduced pressure. The residue was purifiedby flash column chromatography (SiO₂, gradient Hex:EtOAc 4:1 toEtOAc) toafford 108 (21.6 mg, 70%) as a white solid. Rf: 0.58 (Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 7.47–7.44 (m, 2H), 7.35–7.34 (m, 3H), 7.29 (d,J=15.9 Hz, 1H), 6.62 (s, 1H), 5.99 (d, J=1.2 Hz, 1H), 5.93 (d, J=1.2 Hz,1H), 5.05 (d, J=5.7 Hz, 1H), 4.94 (d, J=5.7 Hz, 1H), 4.81 (d, J=11.5 Hz,1H), 4.16–4.11 (m, 3H), 3.34 (brd, J=5.4 Hz, 1H), 3.24 (bs, 3H),3.22–3.20 (m, 2H), 2.94 (dd, J₁=8.1 Hz, J₂=18.0 Hz, 1H), 2.80 (d, J=14.1Hz, 1H), 2.64 (d, J=18.0 Hz, 1H), 2.32 (s, 3H), 2.28 (s, 3H), 2.09 (s,3H), 1.94 (s, 3H), 1.71 (dd, J₁=11.7 Hz, J₂=15.6 Hz, 1H).

ESI-MS m/z: Calcd. for C₃₉H₄₁N₃O₉: 695.7. Found (M+1)⁺: 696.2.

Example 103

To a solution of 105 (16 mg, 0.025 mmol) in CH₂Cl₂ (0.2 mL),trifluoroacetic acid (77 μL, 1 mmol) was added at 0° C. and the reactionmixture was stirred for 3.5 h at 23° C. The reaction was quenched at 0°C. with saturated aqueous sodium bicarbonate (15 mL) and extracted withethyl acetate (2×10 mL). The combined organic layers were dried oversodium sulphate, filtered, and the solvent was eliminated under reducedpressure. The residue was purified by flash column chromatography (SiO₂,Hex:EtOAc 1:1) to afford 109 (12 mg, 81%) as a white solid. Rf: 0.32(Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 6.43 (s, 1H), 5.97 (d, J=1.5 Hz, 1H), 5.91 (d,J=1.5 Hz, 1H), 5.69 (s, 1H), 4.51 (dd, J₁=3.3 Hz, J₂=11.1 Hz, 1H),4.10–4.05 (m, 3H), 3.78–3.77 (m, 1H), 3.75 (s, 3H), 3.33 (d, J=8.1 Hz,1H), 3.22 (dt, J₁=2.7 Hz, J₂=12.0 Hz, 1H), 2.96 (dd, J₁=8.4 Hz, J₂=17.7Hz, 1H), 2.80 (d, J=15.6 Hz, 1H), 2.55 (d, J=18.0 Hz, 1H), 2.33 (s, 3H),2.24 (s, 3H), 2.01 (s, 3H), 1.87–1.66 (m, 3H), 1.37–1.27 (m, 2H), 0.77(t, J=7.5 Hz, 3H).

ESI-MS m/z: Calcd. for C₃₂H₃₁N₃O₈: 591.6. Found (M+1)⁺: 592.8.

Example 104

To a solution of 106 (90 mg, 0.1178 mmol) in CH₂Cl₂ (0.3 mL),trifluoroacetic acid (750 μL, 4.71 mmol) was added at 0° C. and thereaction mixture was stirred for 7 h at 23° C. The reaction was quenchedat 0° C. with saturated aqueous sodium bicarbonate (20 mL) and extractedwith ethyl acetate (2×15 mL). The combined organic layers were driedover sodium sulphate, filtered, and the solvent was eliminated underreduced pressure. The residue was purified by flash columnchromatography (SiO₂, Hex:EtOAc 1:1) to afford 110 (71 mg, 84%) as awhite solid. Rf: 0.6 (Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 7.76 (bs, 1H), 7.62–7.57 (m, 2H), 7.48–7.45(m, 1H), 7.12 (d, J=16.2 Hz, 1H), 6.37 (s, 1H), 6.00 (d, J=16.2 Hz, 1H),5.98 (d, J=1.2 Hz, 1H), 5.92 (d, J=1.2 Hz, 1H), 5.60 (bs, 1H), 4.88 (d,J=10.2 Hz, 1H), 4.14 (bs, 1H), 4.10 (d, J=2.4 Hz, 1H), 4.03 (d, J=2.4Hz, 1H), 3.89 (dd, J₁=2.7 Hz, J₂=11.4 Hz, 1H), 3.32 d, J=8.4 Hz, 1H),3.26–3.21 (m, 4H), 2.91 (dd, J₁=8.1 Hz, J₂=18.0 Hz, 1H), 2.82 (d, J=13.8Hz, 1H) 2.58 (d, J=18.0 Hz, 1H), 2.33 (s, 3H), 2.24 (s, 3H), 2.05 (s,3H), 1.89 (s, 3H), 1.84 (dd, J₁=12.0 Hz, J₂=15.6 Hz, 1H).

ESI-MS m/z: Calcd. for C₃₈H₃₆F₃N₃O₈: 719.7. Found (M+1)⁺: 720.3.

Example 105

To a solution of 107 (20 mg, 0.286 mmol) in CH₂Cl₂ (0.2 mL),trifluoroacetic acid (88 μL, 1.144 mmol) was added at 0° C. and thereaction mixture was stirred for 4 h at 23° C. The reaction was quenchedat 0° C. with saturated aqueous sodium bicarbonate (15 mL) and extractedwith ethyl acetate (2×10 mL). The combined organic layers were driedover sodium sulphate, filtered, and the solvent was eliminated underreduced pressure. The residue was purified by flash columnchromatography (SiO₂, Hex:EtOAc 1:1) to afford 111 (18 mg, 96%) as awhite solid. Rf: 0.39 (Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 7.23–7.16 (m, 3H), 7.06–7.04 (m, 2H), 6.43 (s,1H), 5.96 (d, J=1.5 Hz, 1H), 5.90 (d, J=1.5 Hz, 1H), 6.66 (s, 1H), 4.52(dd, J₁=3.3 Hz, J₂=11.1 Hz, 1H), 4.07 (s, 1H), 4.05 (d, J=3.3 Hz, 1H),4.03 (d, J=2.4 Hz, 1H), 3.76 (dd, J₁=3.6 Hz, J₂=11.1 Hz, 1H), 3.56 (s,3H), 3.31 (d, J=7.5 Hz, 1H), 3.23 (d, J=12.0 Hz, 1H), 2.95 (dd, J₁=8.1Hz, J₂=18.0 Hz, 1H), 2.80 (d, J=15.3 Hz, 1H), 2.63–2.58 (m, 2H), 2.53(d, J=18.0 Hz, 1H), 2.33 (s, 3H), 2.61 (s, 3H), 2.21–2.09 (m, 2H), 2.13(s, 3H), 2.02 (s, 3H), 1.85 (dd, J₁=11.7 Hz, J₂₌115.3 Hz, 1H). ESI-MSm/z: Calcd. for C₃₇H₃₉N₃O₈: 653.7. Found (M+1)⁺: 654.3.

Example 106

To a solution of 108 (14 mg, 0.02 mmol) in CH₂Cl₂ (0.4 mL),trifluoroacetic acid (61.5 μL, 0.8 mmol) was added at 0° C. and thereaction mixture was stirred for 6 h at 23° C. The reaction was quenchedat 0° C. with saturated aqueous sodium bicarbonate (15 mL) and extractedwith ethyl acetate (2×10 mL). The combined organic layers were driedover sodium sulphate, filtered, and the solvent was eliminated underreduced pressure. The residue was purified by flash columnchromatography (SiO₂, Hex:EtOAc 2:1) to afford 112 (12 mg, 92%) as awhite solid. Rf: 0.36 (Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 7.46–7.45 (m, 2H), 7.35–7.20 (m, 4H), 6.38 (s,1H), 6.05 (d, J=15.9 Hz, 1H), 5.98 (d, J=1.2 Hz, 1H), 5.93 (d, J=1.2 Hz,1H), 5.57 (s, 1H), 4.71 (d, J=9.3 Hz, 1H), 4.17–4.13 (m, 2H), 4.08 (d,J=1.9 Hz, 1H), 3.89 (dd, J₁=3.6 Hz, J₂=11.4 Hz, 1H), 3.33 (m, 5H),3.26–3.22 (m, 1H), 2.93 (dd, J₁=9.0 Hz, J₂=17.4 Hz 1H), 2.34 (s 3H),2.25 (s, 3H), 2.05 (s, 3H), 1.97 (s, 3H), 1.81 (dd, J₁=12.0 Hz, J₂=15.6Hz, 1H).

ESI-MS m/z: Calcd. for C₃₇H₃₇N₃O₈: 651. Found (M+1)⁺: 652.2.

Example 107

To a solution of 109 (10 mg, 0.017 mmol) in CH₃CN/H₂O (1.5 mL/1 mL),AgNO₃ (86 mg, 0.5 mmol) was added and the reaction was stirred at 23° C.for 24 h. Then, brine (10 mL) and Aq sat NaHCO₃ (10 mL) were added at 0°C. and the mixture was stirred for 15 min, filtered through a pad ofcelite and washed with CH₂Cl₂ (15 mL). The solution was extracted andthe organic layer was dried over Na₂SO₄, filtered and concentrated invacuo. The residue was purified by flash column chromatography (SiO₂,gradient EtOAc to EtOAc:MeOH 3:1) to afford 113 (7 mg, 71%) as a whitesolid.

Rf: 0.41 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 6.45 (s, 1H), 5.95 (d, J=1.5 Hz, 1H), 5.88 (d,J=1.5 Hz, 1H), 5.65 (bs, 1H), 4.50–4.48 (m, 2H), 4.44 (d, J=2.1 Hz, 1H),3.96 (d, J=3.0 Hz, 1H), 3.76 (s, 3H), 3.74–3.70 (m, 1H), 3.30 (d, J=12.3Hz, 1H), 3.13 (d, J=7.5 Hz, 1H), 2.86 (dd, J₁=5.7 Hz, J₂=18.3 Hz, 1H),2.73 (d, J=14.7 Hz, 1H), 2.48 (d, J=17.7 Hz, 1H), 2.33 (s, 3H), 2.24 (s,3H), 2.17 (s, 3H), 2.00 (s, 3H), 1.86–1.55 (m, 3H), 1.42–1.23 (m, 2H),0.75 (t, J=7.5 Hz, 3H).

ESI-MS m/z: Calcd. for C₃₁H₃₈N₂O₉: 582.6. Found (M−17)⁺: 565.3.

Example 108

To a solution of 110 (42.8 mg, 0.059 mmol) in CH₃CN/H₂O (1.5 mL/1 mL).AgNO₃ (303 mg, 1.78 mmol) was added and the reaction was stirred at 23°C. for 24 h. Then, brine (10 mL) and Aq sat NaHCO₃ (10 mL) were added at0° C. and the mixture was stirred for 15 min, filtered through a pad ofcelite and washed with CH₂Cl₂ (20 mL). The solution was extracted andthe organic layer was dried over Na₂SO₄, filtered and concentrated invacuo. The residue was purified by flash column chromatography (SiO₂,gradient EtOAc to EtOAc:MeOH 5:1) to afford 114 (30 mg, 71%) as a whitesolid.

Rf: 0.30 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 7.75 (bs, 1H), 7.61–7.56 (m, 2H), 7.45–7.42(m, 1H), 7.12 (d, J=16.2 Hz, 1H), 6.38 (s, 1H), 6.02 (d, J=16.2 Hz, 1H),5.97 (d, J=1.5 Hz, 1H), 5.90 (d, J=1.5 Hz, 1H), 5.50 (bs, 1H), 4.87 (bs,1H), 4.56 (m, 1H), 4.45 (bs, 1H), 3.92 (d, J=2.4 Hz, 1H), 3.31 (dt,J₁=3.6 Hz, J₂=12.9 Hz, 1H), 3.21 (bs, 3H), 3.13 (d, J=7.8 Hz, 1H), 2.82(dd, J₁=8.1 Hz, J₂=18.0 Hz, 1H), 2.75 (d, J=14.7 Hz, 1H), 2.49 (d,J=18.0 Hz, 1H), 2.33 (s, 3H), 2.21 (s, 3H), 2.05 (s, 3H), 1.89 (s, 3H),1.78 (dd, J₁=12.0 Hz, J₂=15.6 Hz, 1H).

ESI-MS m/z: Calcd. for C₃₇H₃₇F₃N₂O₉: 710.6. Found (M−17)⁺: 693.2.

Example 109

To a solution of 111 (12 mg, 0.018 mmol) in CH₃CN/H₂O (1.5 mL/1 mL),AgNO₃ (93.5 mg, 0.55 mmol) was added and the reaction was stirred at 23°C. for 24 h. Then, brine (10 mL) and Aq sat NaHCO₃ (10 mL) were added at0° C. and the mixture was stirred for 15 min, filtered through a pad ofcelite and washed with CH₂Cl₂ (15 mL). The solution was extracted andthe organic layer was dried over Na₂SO₄, filtered and concentrated invacuo. The residue was purified by flash column chromatography (SiO₂,gradient EtOAc to EtOAc:MeOH 1:1) to afford 115 (10 mg, 86%) as a whitesolid.

Rf: 0.43 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 7.23–7.14 (m, 3H), 7.05–7.03 (m, 2H), 6.45 (s,1H), 5.93 (d, J=1.2 Hz, 1H), 5.88 (d, J=1.2 Hz, 1H), 5.63 (brd, 1H),4.55–4.49 (m, 2H), 4.43 (d, J=2.7 Hz, 1H), 3.96 (d, J=3.1 Hz, 1H),3.80–3.73 (m, 1H), 3.56 (bs, 3H), 3.32 (dt, J₁=3.3 Hz, J₂=12.6 Hz, 1H),3.13 (d, J=6.0 Hz, 1H), 2.86 (dd, J₁=7.5 Hz, J₂=18.3 Hz, 1H), 2.74 (d,J=14.7 Hz, 1H), 2.61–2.56 (m, 2H), 2.47 (d, J=18.0 Hz, 1H), 2.33 (s,3H), 2.23 (s, 3H), 2.13 (s, 3H), 2.01 (s, 3H), 1.99–1.94 (m, 2H), 1.78(dd, J₁=11.7 Hz, J₂=15.0 Hz, 1H).

ESI-MS m/z: Calcd. for C₃₆H₄₀N₂O₉: 644.7. Found (M−17)⁺: 627.2.

Example 110

To a solution of 112 (12 mg, 0.018 mmol) in CH₃CN/H₂O (1.5 mL/1 mL),AgNO₃ (93 mg, 0.55 mmol) was added and the reaction was stirred at 23°C. for 24 h. Then, brine (10 mL) and Aq sat NaHCO₃ (10 mL) were added at0° C. and the mixture was stirred for 15 min, filtered through a pad ofcelite and washed with CH₂Cl₂ (15 mL). The solution was extracted andthe organic layer was dried over Na₂SO₄, filtered and concentrated invacuo. The residue was purified by flash column chromatography (SiO₂,gradient EtOAc to EtOAc:MeOH 1:1) to afford 116 (8 mg, 70%) as a whitesolid.

Rf: 0.41 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 7.44–7.43 (m, 2H), 7.34–7.27 (m, 4H), 6.39 (s,1H), 6.03 (d, J=15.9 Hz, 1H), 5.96 (d, J=1.5 Hz, 1H), 5.90 (d, J=1.5 Hz,1H), 5.55 (m, 1H), 4.47 (m, 1H), 4.50 (m, 1H), 3.94 (d, J=3.6 Hz, 1H),3.85 (dd, J₁=3.3 Hz, J₂=11.1 Hz, 1H), 3.66 (bs, 3H), 3.34–3.31 (m, 2H),3.13 (d, J=5.1 Hz, 1H), 2.93–2.73 (m, 2H), 2.53 (d, J=18.0 Hz, 1H), 2.33(s, 3H), 2.22 (s, 3H), 2.03 (s, 3H), 1.94–1.82 (m, 1H).

ESI-MS m/z: Calcd. for C₃₆H₃₈N₂O₉: 642.7. Found (M−17)⁺: 625.2.

Example 111

To a solution of 17 (6.28 g, 9.06 mmol) in CH₂Cl₂ (45.3 mL), allylchloroformate (3.85 mL, 36.24 mmol) and pyridine (2.93 mL, 36.24 mmol)were added at 0° C. The reaction mixture was stirred for 16 h at 23° C.and then, the solution was diluted with CH₂Cl₂ (150 mL) and washed with0.1 N HCl (2×100 mL). The organic layer was dried over Na₂SO₄, filtered,and the solvent was eliminated under reduced pressure to give 117 (5.96g, 84%) which was used in following steps with no further purification.

Rf: 0.56 (CH₂Cl₂:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 6.72 (s, 1H), 6.05–5.94 (m, 1H), 6.01 (s, 1H),5.91 (s, 1H), 5.44 (dd, J₁=1.2 Hz, J₂=17.1 Hz, 1H), 5.35 (dd, J₁=1.2 Hz,J₂=10.5 Hz, 1H), 5.34 (m, 1H), 5.10 (d, J=5.7 Hz, 1H), 5.05 (d, J=5.7Hz, 1H), 4.68 (d, J=5.7 Hz, 1H), 4.65 (dt, J₁=1.2 Hz, J₂=6 Hz, 1H), 4.18(brd, J=9 Hz, 2H), 4.04 (bs, 1H), 3.70 (s, 3H), 3.67–3.60 (m, 1H), 3.55(s, 3H), 3.43–3.41 (m, 2H), 3.29–3.25 (m, 2H), 3.00 (dd, J₁=8.7 Hz,J₂=18.3 Hz, 1H), 2.90 (dd, J₁=2.4 Hz, J₂=16.2 Hz, 1H), 2.75 (d, J=18.3Hz, 1H), 2.35 (s, 3H), 2.22 (s, 3H), 2.06 (s, 3H), 1.83 (dd, J₁=11.4 Hz,J₂=15.9 Hz, 1H), 1.39 (s, 9H), 0.73 (d, J=6.9 Hz, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 172.1, 152.8, 148.6, 148.3, 144.6, 140.7,140.6, 131.5, 131.2, 131.1, 130.4, 125.3, 125.0, 123.3, 120.9, 119.1,118.8, 117.6, 112.9, 112.0, 101.6, 99.2, 71.8, 69.0, 68.4, 59.7, 59.2,57.6, 57.3, 56.7, 55.8, 55.2, 41.4, 39.9, 28.2, 26.0, 25.0, 18.6, 15.6,9.0.

ESI-MS m/z: Calcd. for C₄₀H₅₁N₅O₁₁: 777.8. Found (M+1)⁺: 778.3

Example 112

To a solution of 117 (3.96 g, 5.09 mmol) in MeOH (37.4 mL),trimetylchlorosilane (6.5 mL, 50.9 mmol) was added at 0° C. The reactionmixture was stirred for 4 h at 23° C. and then, the solvent waseliminated under reduced pressure. The residue was diluted with EtOAc(70 mL) and washed with a saturated aqueous solution of NaHCO₂ (2×45mL). The organic layer was dried over Na₂SO₄, filtered, and the solventwas eliminated in vacuo to give 118 (2.77 g, 86%) which was used infollowing steps with no further purification.

Rf: 0.61 (Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 6.50 (s, 1H), 6.45 (m, 1H), 6.10–6.03 (m, 1H),6.00 (s, 1H), 5.93 (s, 1H), 5.47 (dd, J1=1.2 Hz, J2=17.1 Hz, 1H), 5.38(dd, J1=1.2 Hz, J2=10.5 Hz, 1H), 4.81–4.64 (m, 2H), 4.10–4.03 (m, 3H),3.75 (s, 3H), 3.70–3.44 (m, 2H), 3.35 (d, J=8.1 Hz, 1H), 3.28 (dt,J1=2.7 Hz, J2=9 Hz, 1H), 2.98 (dd, J1=7.8 Hz, J2=18 Hz, 1H), 2.90 (dd,J1=2.7 Hz, J2=16.2 Hz, 1H), 2.78 (dd, J1=6.9 Hz, J2=14.1 Hz, 1H), 2.63(d, J=18.3 Hz, 1H), 2.30 (s, 3H), 2.25 (s, 3H), 2.04 (s, 3H), 1.88 (dd,J1=13.2 Hz, J2=15.6 Hz, 1H), 0.95 (d, J=6.9 Hz, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 175.8, 152.9, 146.6, 144.6, 142.5, 140.8,140.6, 131.5, 131.3, 128.5, 121.1, 120.8, 118.9, 117.8, 117.0, 113.2,111.9, 101.7, 68.9, 60.6, 59.1, 56.6, 56.4, 55.7, 55.2, 50.5, 41.7,39.4, 26.1, 25.0, 21.0, 15.6, 9.2.

ESI-MS m/z: Calcd. for C₃₃H₃₉N₅O₈: 633.6. Found (M+1)⁺: 634.2.

Example 113

To a solution of 118 (3.52 g, 5.56 mmol) in CH₂Cl₂ (28 mL),phenylisothiocyanate (3.99 mL, 33.36 mmol) was added at 23° C. Thereaction mixture was stirred for 3 and then the solvent was eliminatedunder reduced pressure. The residue was purified by flash columnchromatography to afford 119 (3.5 g, 82%) as a white solid.

Rf: 0.52 (CH₂Cl₂:EtOAc 1:5).

¹H NMR (300 MHz, CDCl₃) δ 7.69 (bs, 1H), 7.49–7.46 (m, 2H), 7.34–7.21(m, 2H), 6.96 (d, J=6.9 Hz, 1H), 6.06–5.97 (m, 1H), 6.03 (s 1H), 5.96(bs, 1H), 5.91 (s, 1H), 5.66 (s, 1H), 5.47 (dd, J1=1.5 Hz, J2=17.1 Hz,1H), 5.37 (dd, J1=1.5 Hz, J2=10.5 Hz, 1H), 5.36 (s, 11H), 4.75–4.70 (m,2H), 4.54–4–49 (m, 1H), 4.14 (d, J=2.4 Hz, 1H), 4.07–4.06 (m, 2H), 3.70(s, 3H), 3.44 (m, 1H), 3.35 (d, J=8.1 Hz, 1H), 3.21 (dt, J1=2.7 Hz,J2=6.6 Hz, 1H), 2.94–2.82 (m, 2H), 2.63 (d, J=18 Hz, 1H), 2.24 (s, 3H),2.06 (s, 3H), 2.06 (s, 3H), 1.90 (dd, J1=11.7 Hz, J2=15.9 Hz, 1H), 0.71(d, J=6.9 Hz, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 178.6, 171.9, 152.8, 146.7, 144.5, 142.6,140.8, 140.5, 136.3, 131.3, 131.0, 129.9, 129.8, 128.9, 126.7, 125.2,124.3, 121.1, 120.6, 118.9, 117.7, 116.5, 112.8, 112.1, 101.6, 68.9,60.5, 58.9, 57.3, 56.1, 55.9, 55.1, 53.3, 41.5, 39.2, 25.9, 24.6, 20.9,15.4, 9.1.

ESI-MS m/z: Calcd. for C₄₀H₄₄N₃O₈S: 768.8. Found (M+1)⁺: 769.3.

Example 114

To a solution of 119 (3.38 g, 4.4 mmol) in MeOH (22 mL),trimetylchlorosilane (2.3 mL, 22 mmol) was added at 0° C. The reactionmixture was stirred for 1.5 h at 23° C. and then, the solvent waseliminated under reduced pressure. The residue was diluted with EtOAc(100 mL) and washed with 0.1 N HCl (2×75 mL). The aqueous phase wasbasified with a saturated aqueous solution of NaHCO₂ and extracted withCH₂Cl₂ (2×100 mL). The combined organic layers were dried over Na₂SO₄,filtered, and the solvent was eliminated under reduced pressure toafford 120 (2.47 g, 100%) as a white solid which was used in followingsteps with no further purification.

Rf: 0.26 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 6.45 (s, 1H), 6.05–5.98 (m, 1H), 5.97 (d,J=1.2 Hz, 1H), 5.90 (d, J=1.2 Hz, 1H), 5.44 (dd, J1=1.2 Hz, J2=17.1 Hz,1H), 5.35 (dd, J1=1.2 Hz, J2=10.2 Hz, 1H), 4.75–4.71 (m, 2H), 4.12–4.10(m, 1H), 3.99 (d, J=2.4 Hz, 1H), 3.92 (bs, 1H), 3.73 (s, 3H), 3.36–3.26(m, 2H), 3.06 (dd, J1=8.4 Hz, J2=18 Hz, 1H), 2.89 (dd, J1=2.7 Hz,J2=15.9 Hz, 1H), 2.75–2.73 (m, 2H), 2.48 (d, J=18 Hz, 1H), 2.32 (s, 3H),2.23 (s, 3H), 2.05 (s, 3H), 1.85 (dd, J1=11.7 Hz, J2=15.6 Hz, 1H).

¹³C NMR (75 MHz, CDCl₃) δ 153.0, 146.6, 144.5, 142.8, 140.7, 131.5,130.5, 128.9, 121.3, 120.9, 119.1, 117.9, 116.7, 113.8, 111.6, 101.5,69.0, 60.6, 59.8, 58.7, 56.5, 56.0, 55.3, 44.2, 41.8, 31.6, 26.1, 25.7,15.7, 9.2.

ESI-MS m/z: Calcd. for C₃₀H₃₄N₄O₇: 562.6. Found (M+1)⁺: 563.2.

Example 115

To a solution of 120 (2.57 g, 4.4 mmol) in CH₂Cl₂ (44 mL), TrocCl (0.91mL, 6.6 mmol) and pyridine (0.53 mL, 6.6 mmol) were added at −20° C. Thereaction mixture was stirred for 30 min at 0° C. and then, the solutionwas diluted with CH₂Cl₂ (50 mL) and washed with 0.1 N HCl (2×25 mL). Theorganic layer was dried over Na₂SO₄, filtered, and the solvent waseliminated under reduced pressure to give 121 (3.24 g, 100%) which wasused in following steps with no further purification.

Rf: 0.62 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 6.50 (s, 1H), 6.07–6.01 (m, 1H), 5.99 (d,J=1.2 Hz, 1H), 5.93 (d, J=1.2 Hz, 1H), 5.68 (s, 1H), 5.46 (dd, J1=1.2Hz, J2=17.1 Hz, 1H), 5.37 (dd, J1=1.2 Hz, J2=10.5 Hz, 1H), 4.74 (t,J=5.7 Hz, 2H), 4.63–4.62 (m, 1H), 4.54 (d, J=12 Hz, 1H), 4.30 (d, J=12Hz, 1H), 4.14–4.11 (m, 2H), 4.02–4.01 (m, 2H), 3.75 (s, 3H),3.36–3.26(m, 3H), 3.04 (dd, J1=8.1 Hz, J2=17.7 Hz, 1H), 2.91 (dd, J1=2.4Hz, J2=15.6 Hz, 1H), 2.60 (d, J=17.7 Hz, 1H), 2.31 (s, 3H), 2.25 (s,3H), 2.04 (s, 3H), 1.84 (dd, J1=12 Hz, J2=15.9 Hz, 1H).

ESI-MS m/z: Calcd. for C₃₃H₃₅Cl₃N₄O₉: 738.0. Found (M+1)⁺: 737.2.

Example 116

To a solution of 121 (0.45 g, 0.60 mmol) in CH₃CN (4 mL),diisopropylethylamine (2.17 mL, 12.46 mmol), bromomethyl methyl ether(0.76 mL, 9.34 mmol) and dimethylaminopyridine (8 mg, 0.062 mmol) wereadded at 0° C. The reaction mixture was heated at 40° C. for 5 h. Then,the reaction was diluted with CH₂Cl₂ (50 mL) and washed with 0.1 N HCl(2×25 mL). The organic layer was dried over Na₂SO₄, filtered, and thesolvent was eliminated under reduced pressure to give 122 (0.453 g, 95%)which was used in following steps with no further purification.

Rf: 0.31 (RP-18 CH₃CN—H₂O 8:2).

¹H NMR (300 MHz, CDCl₃) δ 6.70 (s, 1H), 6.05–5.99 (m, 1H), 5.97 (s, 1H),5.92 (s, 1H), 5.43 (dd, J1=1.2 Hz, J2=17.1 Hz, 1H), 5.34 (dd, J1=1.2 Hz,J2=10.5 Hz, 1H), 5.10–5.04 (m, 2H), 4.72–4.68 (m, 2H), 4.60 (t, J=5.7Hz, 1H), 4.49 (d, J=12.3 Hz, H), 4.38 (d, J=12.3 Hz, 1H), 4.18 (d, J=2.7Hz, 1H), 4.03–4.00 (m, 2H), 3.71 (s, 3H), 3.54 (s, 3H), 3.38–3.22 (m,4H), 3.04 (dd, J1=7.8 Hz, J2=18.3 Hz, 1H), 2.91 (dd, J1=2.4 Hz, J2=15.9Hz, 1H), 2.61 (d, J=18 Hz, 1H), 2.31 (s, 3H), 2.20 (s, 3H), 2.03 (s,3H), 1.76 (dd, J1=11.7 Hz, J2=15.6 Hz, 1H).

ESI-MS m/z: Calcd. for C₃₃H₃₉Cl₃N₄O₁₀: 782.0. Found (M+1)⁺: 783.2.

Example 117

To a suspension of 122 (0.45 g, 0.579 mmol) in 90% aqueous acetic acid(6 mL), powder zinc (0.283 g, 4.34 mmol) was added and the reaction wasstirred for 6 h at 23° C. Then, the mixture was filtered through a padof celite which was washed with CH₂Cl₂ (25 mL). The organic layer waswashed with an aqueous sat. solution of sodium bicarbonate (pH=9) (2×15mL), dried over Na₂SO₄, filtered, and the solvent was eliminated underreduced pressure to give 123 (0.351 g, 100%) which was used in followingsteps with no further purification.

Rf: 0.38 (SiO₂, EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 6.68 (s, 1H), 6.06–5.99 (m, 1H), 5.97 (d,J=1.5 Hz, 1H), 5.91 (d, J=1.25 Hz, 1H), 5.44 (dd, J1=1.5 Hz, J2=17.4 Hz,1H), 5.36 (dd, J1=1.5 Hz, J2=10.2 Hz, 1H), 5.08 (q, J=5.7 Hz, 2H),5.74–4.70 (m, 2H), 4.02 (d, J=3 Hz, 1H), 4.00 (d, J=2.4 Hz, 1H), 3.91(m, 1H), 3.71 (s, 3H), 3.56 (s, 3H), 3.37–3.35 (m, 1H), 3.29 (t, J=2.7Hz, 1H), 3.08 (dd, J1=7.5 Hz, J2=18 Hz, 1H), 2.90 (dd, J1=2.7 Hz,J2=15.9 Hz, 1H), 2.74 (dd, J1=2.4 Hz, J2=5.1 Hz, 2H), 2.48 (d, J=18 Hz,1H), 2.35 (s, 3H), 2.20 (s, 3H), 2.05 (s, 3H), 1.80 (dd, J1=12 Hz,J2=15.9 Hz, 2H).

ESI-MS m/z: Calcd. for C₃₂H₃₈N₄O₈: 606.6. Found (M+1)⁺: 607.3.

Example 118

To a solution of 120 (100 mg 0.177 mmol) in CH₂Cl₂ (0.7 mL) cinnamoylchloride (29.5 mg, 0.177 mmol) and pyridine (14.37 μL, 0.177 mmol) wereadded at 0° C. The reaction mixture was stirred for 1.5 h and then, thesolution was diluted with CH₂Cl₂ (15 mL) and washed with 0.1 N HCl (10mL). The organic layer was dried over Na₂SO₄, filtered, and the solventwas eliminated under reduced pressure. The residue was purified by flashcolumn chromatography (SiO₂, gradient Hex:EtOAc 2:1 to Hex:EtOAc 1:3) toafford 124 (86 mg, 70%) as a white solid.

Rf: 0.77 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 7.39–7.26 (m, 5H), 7.25 (d, J=15.6 Hz, 1H),6.44 (s, 1H), 6.01 (d, J=1.2 Hz, 1H), 5.94 (d, J=1.2 Hz, 1H), 5.68 (s,1H), 5.65 (d, J=15.6 Hz, 1H), 5.44 (dd, J1=1.2 Hz, J2=17.1 Hz, 1H), 5.35(dd, J1=1.2 Hz, J2=10.5 Hz, 1H), 5.18 (t, J=6 Hz, 1H), 4.73–4.69 (m,2H), 4.11–4.09 (m, 3H), 3.66–3.58 (m, 2H), 3.65 (s, 3H), 3.38–3.31 (m,3H), 3.02 (dd, J1=8.4 Hz, J2=18.3 Hz, 1H), 2.92 (dd, J1=2.7 Hz, J2=15.6Hz, 1H), 2.59 (d, J=18.3 Hz, 1H), 2.31 (s, 3H), 2.05 (s, 3H), 2.02 (s,3H), 1.89 (dd, J1=12.3 Hz, J2=16.2 Hz, 1H).

¹³C NMR (75 MHz, CDCl₃) δ 165.5, 152.7, 146.6, 144.4, 142.6, 140.7,140.5, 140.1, 134.7, 131.2, 130.6, 129.3, 128.7, 128.4, 127.6, 120.8,120.5, 120.3, 118.9, 117.6, 116.5, 113.2, 111.8, 101.6, 68.8, 60.4,59.0, 56.2, 56.1, 55.7, 55.0, 41.5, 40.6, 25.9, 25.1, 15.5, 9.0.

ESI-MS m/z: Calcd. for C₃₉H₄₀N₄O₈: 692.7. Found (M+1)⁺: 693.2.

Example 119

To a solution of 124 (495 mg, 0.713 mmol) in CH₂Cl₂ (28 mL), acetic acid(163 μL), Pd(PPh₃)₂Cl₂ (50 mg, 0.0713 mmol) and Bu₃SnH (384 μL, 1.42mmol) were added at 0° C. The reaction mixture was stirred for 2 h at23° C. and then, the solution was poured into a pad of flash column(SiO₂, gradient Hex:EtOAc 1:1 to EtOAc) to afford 125 (435 mg, 100%) asa white solid. Rf: 0.22 (Hex:EtOAc 1:2).

¹H NMR (300 MHz, CDCl₃) δ 7.36–7.33 (m, 5H), 7.28 (d, J=15.9 Hz, 1H),6.45 (s, 1H), 5.90 (s, 1H), 5.83 (s, 1H), 5.55 (d, J=15.6 Hz, 1H), 5.24(t, J=12.9 Hz, 1H), 4.17 (d, J=1.8 Hz, 1H), 4.10–4.07 (m, 2H), 3.72 (s,3H), 3.46–3.32 (m, 3H), 3.14–3.00 (m, 2H), 2.54 (d, J=18 Hz, 1H), 2.32(s, 3H), 2.05 (s, 6H), 1.89 (dd, J₁=12 Hz, J2=15.3 Hz, 1H).

¹³C NMR (75 MHz, CDCl₃) δ 165.7, 146.9, 145.1, 144.2, 143.0, 140.8,136.5, 134.5, 130.6, 129.4, 128.9, 127.9, 127.7, 120.8, 119.8, 117.8,114.1, 112.9, 107.1, 100.8, 60.5, 59.2, 56.4, 56.0, 55.1, 41.4, 30.7,25.5, 25.3, 15.5, 8.9.

ESI-MS m/z: Calcd. for C₃₅H₃₆N₄O₆: 608.6. Found (M+1)⁺: 609.2.

Example 120

To a solution of 125 (86 mg, 0.124 mmol) in CH₃CN/H₂O (1.5 mL/1 mL),AgNO₃ (632 mg, 3.72 mmol) was added and the reaction was stirred at 23°C. for 24 h. Then, brine (10 mL) and Aq sat NaHCO₃ (10 mL) were added at0° C. and the mixture was stirred for 15 min, filtered through a pad ofcelite and washed with CH₂Cl₂ (20 mL). The solution was extracted andthe organic layer was dried over Na₂SO₄, filtered and concentrated invacuo. The residue was purified by flash column chromatography (SiO₂,gradient EtOAc to EtOAc:MeOH 2:1) to afford 126 (70 mg, 83%) as a whitesolid.

Rf: 0.07 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 7.40–7.28 (m, 5H), 7.25 (d, J=15.6 Hz, 1H),6.48 (s, 1H), 6.00–5.94 (m, 1H), 5.96 (s, 1H), 5.92 (s, 1H), 5.89 (s,1H), 5.53 (d, J=15.6 Hz, 1H), 5.42–5.36 (m, 2H), 5.31 (dd, J₁=1.2 Hz,J2=10.8 Hz, 1H), 4.71–4.65 (m, 2H), 4.51 (d, J=3 Hz, 1H), 4.42 (bs, 1H),4.07 (bs, 1H), 3.79 (dd, J₁=6.9 Hz, J2=12.9 Hz, 1H), 3.68 (s, 3H),3.62–3.59 (m, 1H), 3.41–3.37 (m, 1H), 3.16 (d, J=7.8 Hz, 1H), 2.95 (dd,J₁=7.5 Hz, J2=17.4 Hz, 1H), 2.88–2.83 (m, 1H), 2.43 (d, J=18 Hz, 1H),2.28 (s, 3H), 2.10 (s, 3H), 2.00 (s, 3H), 1.81 (dd, J₁=11.7 Hz, J2=15.3Hz, 1H).

¹³C NMR (75 MHz, CDCl₃) δ 165.5, 152.9, 146.7, 144.5, 144.4, 142.7,141.0, 140.0, 134.6, 131.4, 130.7, 129.2, 128.8, 128.5, 127.8, 127.7,124.6, 121.2, 120.9, 118.9, 116.5, 114.9, 114.7, 111.3, 101.6, 93.3,92.3, 83.2, 68.9, 60.6, 57.8, 56.8, 56.6, 56.3, 52.5, 52.2, 41.6, 26.1,24.6, 15.6, 9.1.

ESI-MS m/z: Calcd. for C₃₁H₄₁N₃O₉: 683.7 Found (M−17)⁺: 666.3

Example 121

To a solution of 120 (1.61 g, 2.85 mmol) in CH₂Cl₂ (4 mL),hydrocinnamoyl chloride (423 μL, 2.85 mmol) and pyridine (230 μL, 2.85mmol) were added at 0° C. The reaction mixture was stirred for 1.5 h andthen, the solution was diluted with CH₂Cl₂ (50 mL) and washed with 0.1 NHCl (30 mL). The organic layer was dried over Na₂SO₄, filtered, and thesolvent was eliminated under reduced pressure. The residue was purifiedby flash column chromatography (SiO₂, gradient Hex:EtOAc 2:1 to EtOAc)to afford 127 (1.64 g, 83%) as a white solid.

Rf: 0.63 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 7.26–7–14 (m, 3H), 7.04–7.01 (m, 2H), 6.44 (s,1H), 6.07–5.99 (m, 1H), 5.97 (d, J=1.5 Hz, 1H), 5.91 (d, J=1.5 Hz, 1H),5.75 (bs, 1H), 5.45 (dd, J₁=1.5 Hz, J2=17.4 Hz, 1H), 5.36 (dd, J₁=1.5Hz, J2=10.2 Hz, 1H), 5.03 (t, J=5.7 Hz, 1H), 5.74–5.66 (m, 2H), 4.09 (d,J=2.4 Hz, 1H), 4.01 (bs, 1H), 3.97 (d, J=2.7 Hz, 1H), 3.62 (dd, J₁=8.4Hz, J2=13.5 Hz, 1H), 3.42 (s, 3H), 3.37–3.28 (m, 3H), 3.04–2.87 (m, 3H),2.67–2.46 (m, 4H), 2.29 (s, 3H), 2.05 (s, 3H), 2.03 (s, 3H), 1.83–1.79(m, 1H).

¹³C NMR (75 MHz, CDCl₃) δ 171.8, 152.8, 146.7, 144.5, 144.4, 142.7,140.9, 140.8, 140.6, 131.4, 130.7, 128.9, 128.4, 128.2, 128.1, 126.0,120.8, 120.4, 118.9, 117.6, 116.6, 113.0, 111.9, 101.6, 68.9, 60.3,59.0, 56.3, 56.2, 55.6, 55.1, 41.6, 40.3, 37.7, 31.0, 25.9, 25.2, 15.5,9.1.

ESI-MS m/z: Calcd. for C₃₉H₂N₄O₈: 694.3. Found (M+1)⁺: 695.3.

Example 122

To a solution of 127 (50 mg, 0.072 mmol) in CH₃CN/H₂O (1.5 mL/1 mL),AgNO₃ (444 mg, 2.16 mmol) was added and the reaction was stirred at 23°C. for 24 h. Then brine (10 mL) and Aq sat NaHCO₃ (10 mL) were added at0° C. and the mixture was stirred for 15 min, filtered through a pad ofcelite and washed with CH₂Cl₂ (15 mL). The solution was extracted andthe organic layer was dried over Na₂SO₄, filtered and concentrated invacuo. The residue was purified by flash column chromatography (SiO₂,gradient EtOAc to EtOAc:MeOH 3:1) to afford 128 (30 mg, 61%) as a whitesolid.

Rf: 0.65 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 7.22–7.11 (m, 3H), 7.06–7.03 (m, 2H), 6.43 (s,1H), 6.08–5.98 (m, 1H), 5.96 (d, J=1.5 Hz, 1H), 5.90 (d, J=1.5 Hz, 1H),5.66 (bs, 1H), 5.44 (dd, J₁=1.5 Hz, J2=17.4 Hz, 1H), 5.36 (dd, J₁=1.5Hz, J2=10.5 Hz, 1H), 4.78–4.65 (m, 2H), 4.44 (d, J=3 Hz, 1H), 4.36 (bs,1H), 3.99 (td, J1=2.1 Hz, J2=9.9 Hz, 1H), 3.78–3.67 (m, 1H), 3.56 (dt,J1=1.5 Hz, J2=11.1 Hz, 1H), 3.43 (s, 3H), 3.30–3.12 (m, 2H), 3.02–2.89(m, 1H), 2.83 (dd, J1=2.7 Hz, J2=15.9 Hz, 1H), 2.62–2.51 (m, 2H), 2.36(d, J=18.6 Hz, 1H), 2.27 (s, 3H), 2.02 (s, 3H), 2.00 (s, 3H), 1.86–1.66(m, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 171.6, 146.7, 141.2, 141.1, 131.5, 130.5,128.9, 128.3, 128.2, 128.2, 125.9, 124.7, 121.1, 121.0, 118.8, 111.3,101.6, 94.0, 83.2, 68.8, 60.3, 57.9, 56.6, 56.3, 52.3, 52.0, 41.7, 41.6,41.1, 37.9, 31.1, 31.0, 26.1, 24.6, 15.5, 9.2.

ESI-MS m/z: Calcd. for C₃₈H₄₃N₃O₉: 685.7. Found (M−17)⁺: 668.3.

Example 123

To a solution of 127 (1.64 g, 2.36 mmol) in CH₃CN (12 mL),diisopropylethylamine (8.22 mL, 47.2 mmol), bromomethyl methyl ether(2.89 mL, 35.4 mmol) and dimethylaminopyridine (29 mg, 0.236 mmol) wereadded at 0° C. The reaction mixture was heated at 40° C. for 5 h. Then,the reaction was diluted with CH₂Cl₂ (80 mL) and washed with 0.1 N HCl(3×25 mL). The organic layer was dried over Na₂SO₄, filtered and thesolvent was eliminated under reduced pressure to give 129 (1.46 g, 84%)which was used in following steps with no further purification.

Rf: 0.24 (RP-18 CH₃CN—H₂O 8:2).

¹H NMR (300 MHz, CDCl₃) δ 7.27–7.11 (m, 3H), 7.05–7.02 (m, 2H), 6.67 (s,1H), 6.08–5.98 (m, 1H), 5.96 (d, J=1.2 Hz, 1H), 5.90 (d, J=1.2 Hz, 1H),5.44 (dd, J1=1.2 Hz, J2=17.1 Hz, 1H), 5.34 (dd, J1=1.2 Hz, J2=10.5 Hz,1H), 5.05 (d, J=6 Hz, 1H), 5.00 (d, J=6 Hz, 1H), 4.97 (t, J=5.1 Hz, 1H),4.75–4.68 (m, 2H), 4.16 (d, J=2.7 Hz, 1H), 3.98–3.97 (m, 1H), 3.68–3.67(m, 1H), 3.65–3.61 (m, 1H), 3.52 (s, 3H), 3.35 (s, 3H), 3.32–3.26 (m,3H), 3.05–2.86 (m, 3H), 2.59–2.48 (m, 2H), 2.30 (s, 3H), 2.02 (s, 3H),1.94 (s, 3H), 1.91–1.67 (m, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 171.4, 152.7, 148.5, 148.3, 144.5, 140.9,140.8, 140.4, 131.1, 130.9, 130.4, 130.1, 128.4, 128.2, 126.0, 124.6,123.7, 120.3, 119.0, 112.9, 111.8, 101.6, 99.1, 68.9, 59.4, 59.1, 57.5,56.7, 56.3, 55.4, 55.1, 41.5, 40.2, 37.7, 30.9, 25.8, 25.2, 15.5, 9.0.

ESI-MS m/z: Calcd. for C₄₁H₄₆N₄O₉: 738.8. Found (M+23)⁺: 761.2.

Example 124

To a solution of 129 (1.46 g, 1.97 mmol) in CH₂Cl₂ (40 mL), acetic acid(450 μL), Pd(PPh₃)₂Cl₂ (138 mg, 0.197 mmol) and Bu₃SnH (1.06 mL, 3.95mmol) were added at 0° C. The reaction mixture was stirred for 5 h at23° C. and then, the solution was poured into a pad of flash column(SiO₂, gradient Hex:EtOAc 1:1 to EtOAc) to afford 130 (1.1 g, 85%) as awhite solid. Rf: 0.22 (Hex:EtOAc 1:2).

¹H NMR (300 MHz, CDCl₃) δ 7.21–7.12 (m, 3H), 6.98–6.95 (m, 2H), 5.86 (s1H), 5.84 (s, 1H), 5.79 (bs, 1H), 5.26 (d, J=6 Hz, 1H), 5.11 (d, J=6 Hz,1H), 5.05 (t, J=5.7 Hz, 1H), 4.19 (d, J=2.4 Hz, 1H), 4.03 (d, J=2.4 Hz,1H), 3.99 (bs, 1H), 3.65 (s, 3H), 3.56 (s, 3H), 3.53–3.42 (m, 2H), 3.34(d, J=8.7 Hz, 1H), 3.27 (brd, J=11.7 Hz, 1H), 3.11 (d, J=15 Hz, 1H),2.99 (dd, J1=8.4 Hz, J2=18.3 Hz, 1H), 2.64–2.52 (m, 3H), 2.29 (s, 3H),2.08 (s, 3H), 2.06 (s, 3H), 1.84 (t, J=7.8 Hz, 2H), 1.71 (dd, J1=12.9Hz, J2=13.5 Hz, 1H).

¹³C NMR (75 MHz, CDCl₃) δ 171.7, 149.0, 147.6, 140.6, 132.1, 131.9,130.9, 130.5, 128.5, 128.4, 128.3, 128.0, 126.0, 124.9, 124.6, 123.1,117.6, 100.8, 99.6, 59.6, 58.9, 57.6, 56.6, 56.5, 55.6, 55.1, 41.5,37.8, 31.5, 31.1, 25.9, 25.1, 22.6, 15.5, 8.8.

ESI-MS m/z: Calcd. for C₃₇H₄₂N₄O₇: 654.7. Found (M⁺+Na): 655.1

Example 125

To a solution of 130 (130 mg, 0.198 mmol) in CH₂Cl₂ (1 mL),trifluoroacetyl anhydride (41.9 μL, 0.297 mmol) and pyridine (24 μL,0.297 mmol) were added at 0° C. The reaction mixture was stirred for 2.5h and then, the solution was diluted with CH₂Cl₂ (10 mL) and washed with0.1 N HCl (7 mL). The organic layer was dried over Na₂SO₄ filtered, andthe solvent was eliminated under reduced pressure. The residue waspurified by flash column chromatography (SiO₂, gradient Hex:EtOAc 4:1 toHex:EtOAc 1:4) to afford 131 (93 mg, 62%) as a white solid.

Rf: 0.30 (Hex:EtOAc 1:2).

¹H NMR (300 MHz, CDCl₃) δ 7.25–7.16 (m, 3H), 7.04–7.02 (m, 2H), 6.78 (s,1H), 6.02 (d, J=1.2 Hz, 1H), 5.95 (d, J=1.2 Hz, 1H), 5.11 (d, J=6.6 Hz,1H), 4.98 (d, J=6.6 Hz, 1H), 4.95 (t, J=6.3 Hz, 1H), 4.61 (bs, 1H), 4.30(s, 1H), 4.08 (s, 1H), 3.96 (d, J=7.2 Hz, 1H), 3.66–3.54 (m, 1H), 3.50(s, 3H), 3.39 (s, 3H), 3.19 (dd, J1=7.8 Hz, J2=18.3 Hz, 1H), 2.88 (d,J=18.6 Hz, 1H), 2.79 (dd, J1=2.7 Hz, J2=15.9 Hz, 1H), 2.66–2.62 (m, 1H),2.57 (s, 3H), 2.06 (s, 6H), 1.94–1.87 (m, 1H), 1.77–1.68 (m, 2H).

ESI-MS m/z: Calcd. for C₃₉H₄₁F₃N₄O₈: 750.7. Found (M+Na)⁺: 751.2.

Example 126

To a solution of 130 (130 mg, 0.198 mmol) in CH₂Cl₂ (2 mL), chloroacetylchloride (23.65 μL, 0.297 mmol) and pyridine (24 μL, 0.297 mmol) wereadded at 0° C. The reaction mixture was stirred for 1.5 h and then, thesolution was diluted with CH₂Cl₂ (10 mL) and washed with 0.1 N HCl (7mL). The organic layer was dried over Na₂SO₄, filtered, and the solventwas eliminated under reduced pressure. The residue was purified by flashcolumn chromatography (SiO₂, gradient Hex:EtOAc 2:1 to Hex:EtOAc 1:1) toafford 132 (130 mg, 90%) as a white solid.

Rf: 0.31 (Hex:EtOAc 1:2).

¹H NMR (300 MHz, CDCl₃) δ 7.24–7.15 (m, 3H), 7.07–7.05 (m, 2H), 6.69 (s,1H) 6.00 (d, J=1.5 Hz, 1H), 5.94 (d, J=1.5 Hz, 1H), 5.11 (d, J=5.7 Hz,1H), 5.04 (d, J=5.7 Hz, 1H), 4.93 (m, 1H), 4.36 (s, 2H), 4.16 (d, J=2.7Hz, 1H), 4.01 (m, 2H), 3.64 (dd, J1=6.9 Hz, J2=12.3 Hz, 1H), 3.54 (s,3H), 3.40 (s, 3H), 3.38–3.35 (m, 2H), 2.29 (dt, J1=3 Hz, J2=12 Hz, 1H),3.03 (dd, J1=7.8 Hz, J2=18 Hz, 1H), 2.77 (dd, J1=2.4 Hz, J2=16.2 Hz,1H), 2.58–2.52 (m, 3H), 2.32 (s, 3H), 2.02 (s, 3H), 1.92–1.85 (m, 1H),1.76–1.65 (m, 2H).

¹³C NMR(75 MHz, CDCl₃) δ 171.6, 164.9, 148.3, 144.6, 140.9, 140.8,139.8, 132.1, 131.9, 131.1, 130.0, 128.2, 126.0, 125.0, 124.6, 123.5,120.1, 117.5, 113.0, 111.5, 101.7, 99.1, 64.9, 59.7, 58.9, 57.7, 56.6,56.4, 55.2, 55.1, 41.5, 40.2, 39.9, 37.7, 30.9, 26.3, 25.1, 15.4, 9.1.

ESI-MS m/z: Calcd. for C₃₉H₄₃ClN₄O₈: 730.2. Found (M+1)⁺: 731.1.

Example 127

To a solution of 130 (130 mg, 0.198 mmol) in CH₂Cl₂ (2 mL),chloropropionyl chloride (28.35 μL, 0.297 mmol) and pyridine (24 μL,0.297 mmol) were added at 0° C. The reaction mixture was stirred for 1.5h and then, the solution was diluted with CH₂Cl₂ (10 mL) and washed with0.1 N HCl (7 mL). The organic layer was dried over Na₂SO₄, filtered, andthe solvent was eliminated under reduced pressure. The residue waspurified by flash column chromatography (SiO₂, Hex:EtOAc 1:1) to afford133 (94 mg, 64%) as a white solid.

Rf: 0.43 (Hex:EtOAc 1:2).

¹H NMR (300 MHz, CDCl₃) δ 7.23–7.12 (m, 3H), 7.06–7.04 (m, 2H), 6.69 (s,1H), 5.97 (s, 1H), 5.92 (s, 1H), 5.08 (d, J=6 Hz, 1H), 5.00 (d, J=6 Hz,1H), 4.97 (m, 1H), 4.16 (bs, 1H), 4.00 (m, 1H), 3.88 (t, J=6.9 Hz, 2H),3.75 (t, J=6.9 Hz, 2H), 3.59 (dd, J1=6.3 Hz, J2=12.3 Hz, 1H), 3.53 (s,3H), 3.37 (s, 3H), 3.03–3.26 (m, 1H), 3.17–2.97 (m, 3H), 2.83–2.73 (m,2H), 2.58–2.52 (m, 3H), 2.31 (s, 3H), 2.03 (s, 6H), 1.93–1.86 (m, 1H),1.79–1.64 (m, 2H).

¹³C NMR (75 MHz, CDCl₃) δ 171.9, 167.8, 148.3, 144.7, 140.8, 132.1,132.0, 131.1, 130.2, 128.2, 126.1, 125.2, 124.6, 123.7, 122.2, 120.2,117.6, 114.7, 112.9, 111.8, 101.7, 99.3, 74.9, 65.0, 59.6, 59.0, 57.7,56.7, 56.4, 55.4, 55.1, 41.5, 38.5, 37.8, 37.2, 31.0, 26.4, 25.2, 15.5,9.3.

ESI-MS m/z: Calcd. for C₄₀H₄₅ClN₄O₈: 744.2. Found (M+1)⁺: 745.0.

Example 128

To a solution of 130 (160 mg, 0.244 mmol) in CH₂Cl₂ (2 mL),heptafluorobutyryl chloride (54.5 μL, 0.366 mmol) and pyridine (40 μL,0.49 mmol) were added at 0° C. The reaction mixture was stirred for 2 hand then, the solution was diluted with CH₂Cl, (15 mL) and washed with0.1 N HCl (10 mL). The organic layer was dried over Na₂SO₄, filtered,and the solvent was eliminated under reduced pressure. The residue waspurified by flash column chromatography (SiO₂, gradient Hex:EtOAc 2:1 toHex:EtOAc 1:4) to afford 134 (120 mg, 63%) as a white solid.

Rf: 0.40 (Hex:EtOAc 1:2).

¹H NMR (300 MHz, CDCl₃) δ 7.25–7.16 (m, 3H), 7.04–7.02 (m, 2H), 6.77 (s,1H), 6.02 (d, J=1.5 Hz, 1H), 5.96 (d, J=1.5 Hz, 1H), 5.11 (d, J=6.6 Hz,1H), 4.95 (d, J=6.6 Hz, 1H), 4.94 (m, 1H), 4.58 (m, 1H), 4.25 (bs, 1H),4.06 (bs, 1H), 3.88 (d, J=6.9 Hz, 1H), 3.64 (dd, J1=7.5 Hz, J2=12.9 Hz,1H), 3.55–3.53 (m, 1H), 3.49 (s, 3H), 3.38 (s, 3H), 3.17 (dd, J1=8.1 Hz,J2=18.9 Hz, 1H), 2.85 (d, J=18.3 Hz, 1H), 2.77 (dd, J1=2.7 Hz, J2=16.2Hz, 1H), 2.60–2.57 (m, 3H), 2.56 (s, 3H), 2.06 (s, 3H), 2.03 (s, 3H),1.96–1.88 (m, 1H), 1.79–1.69 (m, 2H).

ESI-MS m/z: Calcd. for C₄₁H₄₁F₇N₄O₈: 850.7. Found (M+1)⁺: 851.3.

Example 129

To a solution of 131 (93 mg, 0.123 mmol) in CH₂Cl₂ (1 mL),trifluoroacetic acid (381 μL, 4.95 mmol) was added at 0° C. and thereaction mixture was stirred for 6 h at 23° C. The reaction was quenchedat 0° C. with saturated aqueous sodium bicarbonate (15 mL) and extractedwith ethyl acetate (2×10 mL). The combined organic layers were driedover sodium sulphate, filtered, and the solvent was eliminated underreduced pressure to give 135 (65 mg, 75%) as a white solid which wasused in following steps with no further purification. Rf: 0.26(Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 7.24–7.15 (m, 3H), 7.04–7.01 (m, 2H), 6.45 (s,1H), 6.03 (d, J=1.5 Hz, 1H), 5.97 (d, J=1.5 Hz, 1H), 5.62 (s, 1H), 4.97(m, 1H), 4.09 (d, J=1.8 Hz, 1H), 4.03 (bs, 1H), 3.99 (d, J=2.4 Hz, 1H),3.73 (dd, J1=7.5 Hz, J2=12 Hz, 1H), 3.38 (s, 3H), 3.34–3.28 (m, 3H),3.05 (dd, J1=8.4 Hz, J2=18.3 Hz, 1H), 2.75 (dd, J1=3.3 Hz, J2=16.5 Hz,1H), 2.60–2.47 (m, 3H), 2.30 (s, 3H), 2.05 (s, 3H), 2.02 (s, 3H),1.91–1.65 (m, 3H).

ESI-MS m/z: Calcd. for C₃₇H₃₇F₃N₄O₇: 706.2. Found (M+1)⁺: 707.2.

Example 130

To a solution of 132 (130 mg, 0.177 mmol) in CH₂Cl₂ (1 mL),trifluoroacetic acid (545 μL, 7.08 mmol) was added at 0° C. and thereaction mixture was stirred for 3.5 h at 23° C. The reaction wasquenched at 0° C. with saturated aqueous sodium bicarbonate (15 mL) andextracted with ethyl acetate (2×10 mL). The combined organic layers weredried over sodium sulphate, filtered, and the solvent was eliminatedunder reduced pressure to give 136 (118 mg, 97%) as a white solid whichwas used in following steps with no further purification. Rf: 0.27(Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 7.23–7.13 (m, 3H), 7.06–7.03 (m, 2H), 6.45 (s,1H), 5.98 (d, J=1.2 Hz, 1H), 5.91 (d, J=1.2 Hz, 1H), 5.04 (t, J=4.5 Hz,1H), 4.37 (bs, 2H), 4.13 (d, J=2.1 Hz, 1H), 4.03 (bs, 2H), 3.68–3.61(dd, J1=7.2 Hz, J2=12.3 Hz, 1H), 3.40 (s 3H), 3.37–3.28 (m, 3H), 3.02(dd, J1=8.4 Hz, J2=18.6 Hz 1H), 2.75 (dd, J1=2.7 Hz, J2=15.9 Hz 1H),2.58–2.50 (m, 3H), 2.32 (s, 3H), 2.01 (s, 6H), 1.94–1.67 (m, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 171.8, 165.0, 146.8, 144.6, 142.9, 141.0,140.9, 139.8, 132.0, 130.3, 129.4, 128.5, 128.3, 126.0, 120.8, 120.1,117.4, 116.1, 113.0, 111.5, 101.7, 60.5, 58.7, 56.3, 56.2, 55.2, 55.0,41.5, 40.4, 39.5, 37.7, 31.0, 29.6, 26.4, 25.3, 15.5, 9.2.

ESI-MS m/z: Calcd. for C₃₇H₃₉ClN₄O₇: 686.2. Found (M+1)⁺: 687.2.

Example 131

To a solution of 133 (94 mg, 0.126 mmol) in CH₂Cl₂ (1 mL),trifluoroacetic acid (385 μL, 5.0 mmol) was added at 0° C. and thereaction mixture was stirred for 2.5 h at 23° C. The reaction wasquenched at 0° C. with saturated aqueous sodium bicarbonate (15 mL) andextracted with ethyl acetate (2×10 mL). The combined organic layers weredried over sodium sulphate, filtered, and the solvent was eliminatedunder reduced pressure to give 137 (118 mg, 97%) as a white solid whichwas used in following steps with no further purification. Rf: 0.24(Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 7.25–7.14 (m, 3H), 7.05–7.03 (m, 2H), 6.44 (s,1H), 5.98 (d, J=1.5 Hz, 1H), 5.92 (d, J=1.5 Hz, 1H), 5.82 (s, 1H), 5.20(t, J=4.8 Hz, 1H), 4.07 (d, J=2.1 Hz, 1H), 5.82 (s, 1H), 5.20 (t, J=4.8Hz, 1H), 4.07 (d, J=2.1 Hz, 1H), 4.01 (bs, 1H), 3.98 (d, J=2.4 Hz, 1H),3.93–3.84 (m, 2H), 3.63 (ddd, J1=1.5 Hz, J2=6.9 Hz, J3=12 Hz, 1H), 3.44(bs, 3H), 3.37–3.26 (m, 3H), 3.11–3.06 (m, 2H), 3.01 (dd, J1=8.4 Hz,J2=18.3 Hz, 1H), 2.80 (brd, J=13.8 Hz, 1H), 2.58–2.47 (m, 3H), 2.29 (s,3H), 2.03 (s, 3H), 2.01 (s, 3H), 1.93–1.68 (m, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 171.7, 168.0, 146.7, 144.6, 142.8, 142.1,141.0, 140.8, 140.1, 130.7, 129.0, 128.2, 126.0, 122.2, 120.9, 116.7,114.7, 113.1, 111.7, 102.3, 101.7, 72.0, 60.4, 59.1, 56.4, 56.3, 55.7,55.2, 41.7, 40.3, 38.8, 37.8, 37.1, 31.0, 26.4, 25.2, 15.5, 9.4.

ESI-MS m/z: Calcd. for C₃₈H₄₁ClN₄O₇: 700.2. Found (M+23)⁺: 723.1.

Example 132

To a solution of 134 (46 mg, 0.054 mmol) in CH₂Cl₂ (1 mL),trifluoroacetic acid (166 μL, 2.16 mmol) was added at 0° C. and thereaction mixture was stirred for 10 h at 23° C. The reaction wasquenched at 0° C. with saturated aqueous sodium bicarbonate (15 mL) andextracted with ethyl acetate (2×10 mL). The combined organic layers weredried over sodium sulphate, filtered, and the solvent was eliminatedunder reduced pressure to give 138 (35 mg, 80%) as a white solid whichwas used in following steps with no further purification. Rf: 0.26(Hex:EtOAc 1:1).

¹H NMR (300 MHz, CDCl₃) δ 7.23–7.12 (m, 3H), 7.04–7.01 (m, 2H), 6.45 (s,1H), 6.03 (d, J=1.5 Hz, 1H), 5.97 (d, J=1.5 Hz, 1H), 5.64 (s, 1H), 4.98(m, 1H), 4.09 (d, J=2.1 Hz, 1H), 4.03 (bs, 1H), 3.98 (d, J=2.4 Hz, 1H),3.75 (dd, J1=9.6 Hz, J2=14.1 Hz, 1H), 3.35 (s, 3H), 3.29–3.24 (m, 3H),3.04 (dd, J1=7.8 Hz, J2=18.0 Hz, 1H), 2.74 (dd, J1=3.0 Hz, J2=16.8 Hz,1H), 2.57–2.45 (m, 3H), 2.30 (s, 3H), 2.03 (s, 6H), 1.92–1.64 (m, 3H).ESI-MS m/z: Calcd. for C₃₉H₃₇F₇N₄O₇: 806.7. Found (M+1)⁺: 807.3.

Example 133

To a solution of 136 (45 mg, 0.065 mmol) in CH₂Cl₂ (0.3 mL), acetylchloride (4.65 μL, 0.065 mmol), and pyridine (5.2 μL, 0.065 mmol) wereadded at 0° C. The reaction mixture was stirred for 4 h and then, thesolution was diluted with CH₂Cl₂ (15 mL) and washed with 0.1 N HCl (7mL). The organic layer was dried over sodium sulphate, filtered, and thesolvent was eliminated under reduced pressure. The residue was purifiedby, flash column chromatography (SiO₂, gradient Hex:EtOAc 5:1 to EtOAc)to afford 139 (27 mg, 57%) as a white solid.

Rf: 0.36 (Hex:EtOAc 1:2).

¹H NMR (300 MHz, CDCl₃) δ 7.26–7.14 (m, 3H), 7.07–7.04 (m, 2H), 6.84 (s,1H), 6.00 (d, J=1.2 Hz, 1H), 5.94 (d, J=1.2 Hz, 1H), 4.94 (t, J=5.1 Hz,1H), 4.39–4.38 (m, 2H), 4.02 (bs, 2H), 3.67 (d, J=3 Hz, 1H), 3.60–3.54(m, 1H), 3.47–3.35 (m, 3H), 3.42 (s, 3H), 3.26 (dt, J₁=4.8 Hz, J₂=8.7 Hz1H), 3.02 (dd, J₁=8.1 Hz, J₂=18.3 Hz, 1H), 2.64–2.38 (m, 3H), 2.35 (s,3H), 2.25 (s, 3H), 2.06 (s, 3H), 2.03 (s, 3H), 1.95–1.69 (m, 3H).

ESI-MS m/z: Calcd. for C₃₉H₄₁ClN₄O₈: 729.2. Found (M+23)⁺: 752.3.

Example 134

To a solution of 2 (15 mg, 0.0273 mmol) in CH₂Cl₂ (0.2 mL), acetylchloride (1.94 μL, 0.0273 mmol), and pyridine (2.20 mL, 0.0273 mmol)were added at 0° C. The reaction mixture was stirred for 20 minutes andthen, the solution was diluted with CH₂Cl₂ (15 mL) and washed with 0.1 NHCl (5 mL). The organic layer was dried over sodium sulphate, filtered,and the solvent was eliminated under reduced pressure. The residue waspurified by flash column chromatography (SiO₂, gradient EtOAc toEtOAcMeOH 5:1) to afford 140 (9 mg, 56%) as a light yellow solid. Rf:0.56 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 6.52 (s, 1H), 6.40 (s, 1H), 5.73 (d, J=7.5 Hz,1H), 4.95 (d, J=6.9 Hz, 1H), 4.20 (d, J=1.5 Hz, 1H), 4.00 (s, 3H), 3.86(d, J=4.5 Hz, 1H), 3.79 (s, 3H), 3.78–3.77 (m, 1H), 3.40–3.35 (m, 2H),3.24 (dt, J₁=3.6 Hz, J₂=11.4 Hz, 1H), 3.17 (d, J=7.8 Hz, 1H), 3.11 (d,J=7.5 Hz, 1H), 3.04 (dd, J₁=3.6 Hz, J₂=18.6 Hz, 1H), 2.92 (dt, J₁=3.3Hz, J₂=14.1 Hz, 1H), 2.43 (d, J=18.0 Hz, 1H), 2.37 (s, 3H), 2.29 (s,3H), 1.89 (s, 3H), 1.79 (s, 3H), 1.75 (dd, J₁=2.7 Hz, J₂=6.9 Hz, 1H),0.99 (d, J=7.5 Hz, 3H).

ESI-MS m/z: Calcd. for C₃₁H₃₇N₅O₇: 591.6. Found (M+1)⁺: 592.3.

To a solution of 2 (15 mg, 0.0273 mmol) in CH₂Cl₂ (0.2 mL),trifluoroacetyl anhydride (3.85 μL, 0.0273 mmol was added at 23° C. Thereaction mixture was stirred for 30 minutes and then, the solution wasdiluted with CH₂Cl₂ (15 mL) and washed with 0.1 N HCl (5 mL). Theorganic layer was dried over sodium sulphate, filtered, and the solventwas eliminated under reduced pressure. The residue was purified by flashcolumn chromatography (SiO₂, gradient EtOAc to EtOAcMeOH 4:1) to afford141 (12.1 mg, 69%) as a light yellow solid. Rf: 0.73 (EtOAc:MeOH 5:1).

¹H NMR (300 MHz, CDCl₃) δ 6.90 (d, J=6.6 Hz, 1H), 6.56 (s, 1H), 5.11 (d,J=6.6 Hz, 1H), 4.47 (bs, 1H), 4.23 (bs, 1H), 3.97 (s, 3H), 3.93 (bs,1H), 3.85–3.81 (m, 1H), 3.77 (s, 3H), 3.40–3–36 (m, 2H), 3.23 (dd,J₁=7.2 Hz, J₂=18.6 Hz, 1H), 3.13–3.08 (m, 3H), 1.86 (s, 3H), 1.74 (dd,J₁=10.8 Hz, J₂=16.8 Hz, 1H), 1.07 (d, J=6.9 Hz, 3H).

ESI-MS m/z: Calcd. for C₃₁H₃₄F₃N₅O₇: 645.6. Found (M+1)⁺: 646.3.

Example 136

To a solution of 45 (30 mg, 0.058 mmol) in CH₂Cl₂ (0.87 mL), DIPEA (15.0mL, 0.086 mmol), EDC.HCl (27.6 mg, 0.145 mmol), N-Boc-Phenylalanine(22.9 mg, 0.086 mmol) and DMAP (0.7 mg, 0.006 mmol) were added at roomtemperature and the reaction mixture was stirred for 4 h. Then, thesolution was diluted with CH₂Cl₂ (10 mL) and washed successively with0.1 N HCl (5 mL) and a solution of 10% NaHCO₃ (5 ml). The organic layerwas dried over Na₂SO₄, filtered, and the solvent was eliminated underreduced pressure. The residue was purified by flash columnchromatography (SiO₂, Hex: EtOAc 1:2) to afford 174 (17 mg, 38%) as awhite solid.

Rf=0.35 Hex:AcOEt 1:2.

¹H NMR (300 MHz, CDCl₃) 7.24–7.15 (m, 3H), 7.05–7.02 (m, 2H), 6.43 (s,1H), 5.88 (s, 1H), 5.78 (s, 1H), 5.64 (s, 1H), 5.63 (bs, 1H), 4.80 (bs,1H), 3.98 (s, 1H), 3.85 (bs, 2H), 3.75 (bs, 1H), 3.58 (bs, 1H), 3.53(bs, 3H), 3.38 (m, 1H), 3.17–3.10 (m, 3H) 2.90 (dd, J₁=8.7 Hz, J₂=17.7Hz, 1H), 2.73 (d, J=14.4 Hz, 1H), 2.57 (m, 1H), 2.43–2.37 (m, 1H), 2.25(s, 3H), 2.24 (s, 3H), 2.10 (s, 3H), 1.94 (s, 3H), 1.76 (dd, J₁=12.3 Hz,J₂=15.6 Hz, 1H), 1.19 (bs, 9H). ¹³C NMR (75 MHz, CDCl₃) 171.2, 168.8,146.6, 144.6, 142.8, 140.6, 137.0, 130.7, 129.5, 129.0, 128.4, 126.8,121.1, 121.0, 117.8, 116.7, 113.3, 111.8, 101.5, 60.5, 59.7, 57.0, 56.4,55.3, 41.9, 41.6, 38.7, 31.6, 29.7, 28.2, 26.5, 25.2, 22.6, 20.3, 15.7,14.1, 9.3.

ESI-MS m/z: Calcd. for C₄₂H₄₉N₅O₉: 767.87. Found (M+1)⁺: 768.3.

Example 137

To a solution of 45 (30 mg, 0.058 mmol) in CH₂Cl₂ (0.87 mL), DIPEA (15.0mL, 0.086 mmol), EDC.HCl (27.6 mg, 0.145 mmol), N-Boc-Valine (18.8 mg,0.086 mmol) and DMAP (0.7 mg, 0.006 mmol) were added at room temperatureand the reaction mixture was stirred for 4 h. Then, the solution wasdiluted with CH₂Cl₂ (10 mL) and washed successively with 0.1 N HCl (5mL) and a solution of 10% NaHCO₃ (5 ml). The organic layer was driedover Na₂SO₄, filtered, and the solvent was eliminated under reducedpressure. The residue was purified by flash column chromatography (SiO₂.Hex: EtOAc 1:2) to afford 175 (18 mg, 43%) as a white solid.

Rf=0.25 Hex:EtOAc 1:1.

¹H NMR (300 MHz, CDCl₃) δ 6.42 (s, 1H), 5.97 (s, 1H), 5.82 (s, 1H), 5.73(bs, 1H), 5.50 (bs, 1H), 4.82 (bs, 1H), 4.15 (bs, 1H), 4.03 (bs, 1H),3.96 (bs, 1H), 3.72 (s, 3H), 3.61 (m, 1H), 3.41–3.15 (m, 3H), 2.96 (dd,J₁=8.4 Hz, J₂=18.3 Hz, 1H), 2.72 (d, J=16.5 Hz, 1H), 2.53 (d, J=18 Hz,1H), 2.25 (s, 3H), 2.21 (s, 3H), 1.93 (s, 3H), 1.81 (dd, J₁=14.1 Hz,J₂=14.7 Hz, 1H), 1.34 (s, 9H), 0.83–0.76 (m, 2H), 0.61 (d, J=6.3 Hz,3H), 0.54 (d, J=6.3 Hz, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 171.6, 168.7, 155.4, 146.8, 144.5, 142.9,140.7, 130.7, 128.8, 121.0, 120.6, 117.7, 116.8, 113.3, 111.9, 101.4,60.6, 60.0, 59.3, 57.2, 56.3, 55.2, 41.7, 29.7, 29.3, 28.2, 26.2, 25.2,22.6, 20.3, 18.9, 17.7, 15.7, 14.1, 9.3.

ESI-MS m/z: Calcd. for C₃₈H₄₉N₅O₉: 719.82. Found (M+1)⁺: 720.3.

Example 138

To a solution of 45 (38 mg, 0.073 mmol) in CH₂Cl₂ (1.09 mL), DIPEA (19.0mL, 0.109 mmol), EDC.HCl (34.9 mg, 0.182 mmol), N-Boc-Proline (23.5 mg,0.109 mmol) and DMAP (0.8 mg, 0.007 mmol) were added at 23° C. and thereaction mixture was stirred for 4.5 h. Then, the solution was dilutedwith CH₂Cl₂ (10 mL) and washed successively with 0.1 N HCl (5 mL) and asolution of 10% NaHCO₃ (5 ml). The organic layer was dried over Na₂SO₄,filtered, and the solvent was eliminated under reduced pressure. Theresidue was purified by flash column chromatography (SiO₂, Hex: EtOAc1:1) to afford 176 (33 mg, 63%) as a white solid.

Rf=0.14 Hex:EtOAc 1:2.

¹H NMR (300 MHz, CDCl₃) δ 6.49 (s, 1H), 6.02 (bs, 1H), 5.90 (s, 1H),5.74 (s, 1H), 4.19 (bs, 1H), 4.09 (bs, 1H), 3.98 (bs, 1H), 3.76 (s, 3H),3.38 (d, J=6 Hz, 2H), 3.22 (d, J=11.7 Hz, 1H), 3.15–2.99 (m, 2H), 2.80(d, J=15.3 Hz, 1H), 2.63–2.58 (m, 1H), 2.32 (s, 3H), 2.26 (s, 6H), 1.99(s, 3H), 1.78–1.62 (m, 1H), 1.50–0.83 (m, 7H), 1.21 (s, 9H).

ESI-MS m/z: Calcd. for C₃₈H₄₇N₅O₉: 717.81. Found (M+1)⁺: 718.3.

Example 139

To a solution of 45 (50 mg, 0.144 mmol) in CH₂Cl₂ (0.96 mL), DIPEA (41.8mL, 0.240 mmol), EDC.HCl (46.0 mg, 0.240 mmol), N-Boc-Argininehidrochloride hydrate (47.2 mg, 0.144 mmol) and DMAP (1.1 mg, 0.01 mmol)were added at 23° C. and the reaction mixture was stirred for 4 h. Then,the solvent was removed under vacuum and the residue was purified byflash column chromatography (SiO₂, Hex: EtOAc 1:2) to afford 177 (58 mg,78%) as a white solid.

Rf=0.40 MeOH:EtOAc 1:5.

¹H NMR (300 MHz, CDCl₃) δ 7.53 (bs, 1H), 6.95 (bs, 3H), 6.54 (bs, 1H),6.48 (s, 1H), 6.07 (s, 1H), 6.00 (bs, 1H), 5.88 (s, 1H), 5.11 (bs, 1H),4.23 (s, 1H), 4.08 (s, 1H), 4.02 (s, 1H), 3.76 (s, 3H), 3.70 (bs, 1H),3.48 (bs, 1H), 3.37 (d, J=6.9 Hz, 1H), 3.18 (d, J=10.2 Hz, 1H),3.00–2.94 (m, 3H), 2.82–2.70 (m, 2H), 2.34 (s, 3H), 2.25 (s, 6H), 1.99(s, 3H), 1.73 (brt, J=14.1 Hz, 1H), 1.40 (s, 9H), 1.25 (bs, 3H),0.95–0.85 (m, 2H).

ESI-MS m/z: Calcd. for C₃₉H₅₂N₈O₉: 776.88. Found (M+1)⁺: 777.3.

Example 140

To a solution of 45 (50 mg, 0.096 mmol) in CH₂Cl₂ (1.44 mL), DIPEA (25.8mL, 0.144 mmol), EDC.HCl (46.0 mg, 0.240 mmol), N-Boc-Tryptophan (43.8mg, 0.144 mmol) and DMAP (1.2 mg, 0.009 mmol) were added at 23° C. andthe reaction mixture was stirred for 4 h. Then, the solution was dilutedwith CH₂Cl₂ (10 mL) and washed successively with 0.1 N HCl (5 mL) and asolution of 10% NaHCO₃ (5 ml). The organic layer was dried over Na₂SO₄,filtered, and the solvent was eliminated under reduced pressure. Theresidue was purified by flash column chromatography (SiO₂, Hex: EtOAc1:2) to afford 178 (57 mg, 74%) as a white solid.

Rf=0.12 Hex:EtOAc 1:1.

¹H NMR (300 MHz, CDCl₃) δ 8.50 (bs, 1H), 7.73–7.71 (m, 1H), 7.13–7.12(m, 3H), 6.51 (s, 11H), 5.72 (s, 1H), 5.36 (bs, 1H), 5.28 (bs, 1H), 4.95(bs, 1H), 4.41 (bs, 1H), 4.05 (s, 1H), 3.70 (s, 3H), 3.50 (bs, 2H),3.30–3.17 (m, 4H), 2.89–2.82 (m, 3H), 2.40 (s, 3H), 2.29 (s, 3H), 2.19(s, 3H), 2.03 (s, 3H), 1.49 (s, 9H), 1.26–1.25 (m, 2H).

ESI-MS m/z: Calcd. for C₄₄H₅₀N₆O₉: 806.90. Found (M+1)⁺: 807.3.

Example 141

To a solution of 178 (43 mg, 0.053 mmol) in CH₃CN/H₂O (3 mL/2 mL), AgNO₃(271 mg, 1.60 mmol) was added and the reaction was stirred at 23° C. for17 h. Then, Aq sat NaCl (10 mL) and Aq sat NaHCO₃ (10 mL) were added at0° C. and the mixture was stirred for 15 min, filtered through a pad ofcelite and washed with CH₂Cl, (20 mL). The solution was decanted and theorganic layer was dried and concentrated in vacuo. The residue waspurified by flash column chromatography (SiO₂, EtOAc:MeOH 5:1) to afford179 (24 mg, 56%) as a white solid.

Rf=0.38 EtOAc:MeOH 5:1.

¹H NMR (300 MHz, CDCl₃) δ 8.40 (s, 1H), 7.66 (bs, 1H), 7.25–7.21 (m,1H), 7.16–7.09 (m, 2H), 6.45 (s, 1H), 5.75 (bs, 1H), 5.55 (bs, 1H), 5.45(s, 1H), 5.25 (bs, 1H), 4.36 (bs, 1H), 4.16 (bs, 1H), 4.05 (bs, 1H),3.95 (s, 1H), 3.69 (s, 3H), 3.35–3.02 (m, 6H), 2.83–2.73 (m, 3H), 2.35(s, 3H), 2.24 (s, 3H), 2.19 (s, 3H), 1.99 (s, 3H), 1.77 (dd, J₁=12 Hz,J₂=15.3 Hz 1H).

ESI-MS m/z: Calcd. for C₄₃H₅₁N₅O₁₀: 797.89. Found (M−17)⁺: 780.

Example 142

To a solution of 45 (50 mg, 0.0960 mmol) in CH₂Cl₂ (0.7 mL),2-Chloronicotinoyl chloride (17.7 mg, 0.101 mmol) and pyridine (8.1 mL,0.101 mmol) were added at 0° C. The reaction mixture was stirred for 1.5h and then, the solution was diluted with CH₂Cl₂ (5 mL) and washed with0.1 N HCl (3 mL). The organic layer was dried over Na₂SO₄ filtered, andthe solvent was eliminated under reduced pressure. The residue waspurified by flash column chromatography (SiO₂, Hex: EtOAc 1:1) to afford180 (45 mg, 71%) as a white solid.

Rf=0.18 Hex:EtOAc 1:2.

¹H NMR (300 MHz, CDCl₃) δ 8.32–8.29 (m, 1H), 7.38–7.34 (m, 1H),7.14–7.09 (m, 1H), 6.14 (s, 1H), 5.97 (d, J=1.2 Hz, 1H), 5.92–5.91 (m,2H), 5.75 (d, J=2.1 Hz, 1H), 4.18 (d, J=2.1 Hz, 1H), 4.15 (s, 1H), 4.07(s, 1H), 3.91–3.73 (m, 2H), 3.68 (s, 3H), 3.36 (d, J=7.5 Hz, 1H), 3.31(dt, J₁=2.4 Hz, J₂=11.7 Hz, 1H), 2.92 (dd, J₁=8.1 Hz, J₂=18 Hz, 1H),2.80 (d, J=16.2 Hz, 1H), 2.58 (d, J=18 Hz, 1H), 2.31 (s, 3H), 2.27 (s,3H), 1.99 (s, 3H), 1.91 (s, 3H) 1.97–1.83 (m, 1H).

¹³C NMR (75 MHz, CDCl₃) δ 168.6, 164.8, 150.3, 147.2, 146.5, 144.6,142.5, 140.6, 139.0, 130.9, 130.5, 128.8, 122.3, 120.8, 120.3, 117.6,116.3, 112.7, 112.1, 101.6, 60.6, 58.8, 56.5, 56.3, 55.6, 55.1, 41.6,39.8, 31.5, 26.2, 24.9, 20.3, 15.5, 9.3.

ESI-MS m/z: Calcd. for C₃₄H₃₄ClN₅O₇: 659.2. Found (M+1)⁺: 660.1.

Example 143

To a solution of 180 (39 mg, 0.059 mmol) in CH₃CN/H₂O (3 mL/2 mL), AgNO₃(301 mg, 1.77 mmol) was added and the reaction was stirred at 23° C. for17 h. Then, Aq sat NaCl (10 mL) and Aq sat NaHCO₃ (10 mL) solutions wereadded at 0° C. and the mixture was stirred for 15 min, filtered througha pad of celite and washed with CH₂Cl₂ (20 mL). The solution wasdecanted and the organic layer was dried and concentrated in vacuo. Theresidue was purified by flash column chromatography (SiO₂, EtOAc:MeOH5:1) to afford 181 (28 mg, 73%) as a white solid.

Rf=0.24, EtOAc:MeOH 5:1.

¹H NMR (300 MHz, CDCl₃) δ 8.33–8.31 (m, 1H), 7.40–7.35 (m, 1H),7.16–7.09 (m, 2H), 6.20 (s, 1H), 5.98 (d, J=1.2 Hz, 1H), 5.96 (s, 1H),5.92 (d, J=1.2 Hz, 1H), 5.63 (bs, 1H), 4.60 (bs, 1H), 4.47 (bs, 1H),4.02–3.95 (m, 2H), 3.69 (s, 3H), 3.65–3.56 (m, 1H), 3.48 (s, 3H),3.43–3.38 (m, 1H), 3.17 (brd, J=7.2 Hz, 1H), 2.88 (dd, J₁=8.7 Hz,J₂=18.3 Hz, 1H), 2.74 (d, J=15.3 Hz, 1H), 2.40 (d, J=18.3 Hz, 1H), 2.32(s, 3H), 2.26 (s, 3H), 2.00 (s, 3H), 1.99 (s, 3H), 1.77 (dd, J₁=12 Hz,J₂=15 Hz, 1H).

¹³C NMR (75 MHz, CDCl₃) δ 168.1, 165.0, 150.0, 147.2, 146.5, 144.4,142.5, 140.9, 138.7, 131.5, 130.2, 128.9, 122.3, 121.1, 120.7, 116.1,114.4, 111.4, 101.5, 82.6, 60.6, 57.8, 56.2, 52.1, 41.6, 31.5, 26.4,24.5, 22.6, 20.3, 15.6, 14.1, 9.3.

ESI-MS m/z: Calcd. for C₃₃H₃₅ClN₄O₈: 650.2 Found (M−17)⁺: 633.3.

Example 144

To a solution of 45 (30 mg, 0.058 mmol) in CH₂Cl₂ (0.87 mL). DIPEA (15.0mL, 0.086 mmol), EDC.HCl (27.6 mg, 0.145 mmol), cyclohexylacetic acid(12.2 mg, 0.086 mmol) and DMAP (0.7 mg, 0.006 mmol) were added at 0° C.and the reaction mixture was stirred for 5 h. Then, the solution wasdiluted with CH₂Cl₂ (10 mL) and washed successively with 0.1 N HCl (5mL) and a solution of 10% NaHCO₃ (5 ml). The organic layer was driedover Na₂SO₄, filtered, and the solvent was eliminated under reducedpressure. The residue was purified by flash column chromatography (SiO₂,Hex: EtOAc 1:2) to afford 182 (10 mg, 27%) as a white solid.

Rf=0.11 Hex:EtOAc 1:1.

¹H NMR (300 MHz, CDCl₃) δ 6.50 (s, 1H), 5.98 (d, J=1.2 Hz, 1H), 5.91 (d,J=1.2 Hz, 1H), 5.75 (s, 1H), 5.02–4.91 (m, 1H), 4.11 (bs, 1H), 4.04 (d,J=2.1 Hz, 1H), 4.01 (bs, 1H), 3.78 (s, 3H), 3.72–3.69 (m, 1H), 3.38–3.29(m, 3H), 3.05 (dd, J₁=7.8 Hz, J₂=18.0 Hz, 1H), 2.77 (d, J=15.6 Hz, 1H),2.54 (d, J=18.6 Hz, 1H), 2.33 (s, 3H), 2.32 (s, 3H), 2.27 (s, 3H), 1.98(s, 3H), 1.79 (dd, J₁=11.7 Hz, J₂=15.6 Hz, 1H), 1.59–0.61 (m, 13H).

ESI-MS m/z: Calcd. for C₃₆H₄₄N₄O₇: 644.76. Found (M+1)⁺: 645.3.

Example 145

To a solution of 45 (30 mg, 0.058 mmol) in CH₂Cl₂ (0.87 mL), DIPEA (15.0mL, 0.086 mmol), EDC.HCl (27.6 mg, 0.145 mmol), cyclohexylacetic acid(12.2 mg, 0.086 mmol) and DMAP (0.7 mg, 0.006 mmol) were added at 0° C.and the reaction mixture was stirred for 5 h. Then, the solution wasdiluted with CH₂Cl₂ (10 mL) and washed successively with 0.1 N HCl (5mL) and a solution of 10% NaHCO₃ (5 ml). The organic layer was driedover Na₂SO₄, filtered, and the solvent was eliminated under reducedpressure. The residue was purified by flash column chromatography (SiO₂,Hex: EtOAc 1:2) to afford 183 (17 mg, 38%) as a white solid.

Rf=0.13 Hex:EtOAc 1:1.

¹H NMR (300 MHz, CDCl₃) δ 6.87 (s, 1H), 5.99 (d, J=1.2 Hz, 1H), 5.92 (d,J=1.2 Hz, 1H), 4.95 (t, J=5.7 Hz, 1H), 4.08 (bs, 1H), 4.00 (bs, 1H),3.71 (s, 3H), 3.64 (d, J=1.8 Hz, 2H), 3.38 (d, J=6.6 Hz, 1H), 3.33–3.32(m, 1H), 3.27 (d, J=11.7 Hz, 1H), 3.06 (dd, J₁=7.8 Hz, J₂=18.0 Hz, 1H),2.65–2.59 (m, 1H), 2.50–2.47 (m, 1H), 2.35 (s, 3H), 2.27 (s, 6H), 1.99(s, 3H), 1.78–1.74 (m, 1H) 1.60–0.62 (m, 26H).

ESI-MS m/z: Calcd. for C₄₄H₅₆N₄O₈: 768.94. Found (M+1)⁺: 769.3.

Example 146

To a solution of 45 (30 mg, 0.058 mmol) in CH₂Cl₂ (0.87 mL), DIPEA (15.0mL, 0.086 mmol), EDC.HCl (27.6 mg, 0.145 mmol), cyclohexylpropionic acid(13.5 mg, 0.086 mmol) and DMAP (0.7 mg, 0.006 mmol) were added at 0° C.and the reaction mixture was stirred at 23° C. for 6 h. Then, thesolution was diluted with CH₂Cl₂ (10 mL) and washed successively with0.1 N HCl (5 mL) and a solution of 10% NaHCO₃ (5 ml). The organic layerwas dried over Na₂SO₄, filtered, and the solvent was eliminated underreduced pressure. The residue was purified by flash columnchromatography (SiO₂, Hex: EtOAc 1:2) to afford 184 (15 mg, 39%) as awhite solid.

Rf=0.15 Hex:EtOAc 1:1.

¹H NMR (300 MHz, CDCl₃) δ 6.50 (s, 1H), 5.98 (s, 1H), 5.91 (s, 1H), 5.74(s, 1H), 5.01 (t, J=5.1 Hz, 1H), 4.09 (bs, 1H), 4.06 (s, 1H), 4.02 (bs,1H), 3.76 (s, 3H), 3.64–3.58 (m, 1H), 3.42–3.41 (m, 1H), 3.36 (d, J=7.5Hz, 1H), 3.28 (d, J=12.3 Hz, 1H), 3.05 (dd, J₁=8.6 Hz, J₂=18 Hz, 1H),2.79 (d, J=14.7 Hz, 1H), 2.57 (d, J=18 Hz, 1H), 2.32 (s, 3H), 2.30 (s,3H), 2.25 (s, 3H), 1.99 (s, 3H), 1.77 (dd, J₁=12.0 Hz, J₂=15.9 Hz, 1H),1.62–0.71 (m, 15H). ESI-MS m/z: Calcd. for C₃₇H₄₆N₄O₇: 658.78. Found(M+1)⁺: 659.3.

Example 147

To a solution of 45 (30 mg, 0.058 mmol) in CH₂Cl₂ (0.87 mL), DIPEA (15.0mL, 0.086 mmol), EDC.HCl (27.6 mg, 0.145 mmol), cyclohexylpropionic acid(13.5 mg, 0.086 mmol) and DMAP (0.7 mg, 0.006 mmol) were added at 0° C.and the reaction mixture was stirred for 6 h. Then, the solution wasdiluted with CH₂Cl₂ (10 mL) and washed successively with 0.1 N HCl (5mL) and a solution of 10% NaHCO₃ (5 ml). The organic layer was driedover Na₂SO₄, filtered, and the solvent was eliminated under reducedpressure. The residue was purified by flash column chromatography (SiO₂,Hex: EtOAc 1:2) to afford 185 (21 mg, 46%) as a white solid.

Rf=0.17 Hex:EtOAc 1:1.

¹H NMR (300 MHz, CDCl₃) δ 6.86 (s, 1H), 5.99 (s, 1H), 5.92 (s, 1H), 4.97(t, J=5.4 Hz, 1H), 4.10 (d, J=2.4 Hz, 1H), 4.01 (bs, 1H), 3.70 (s, 3H),3.64 (d, J=2.4 Hz, 1H), 3.51 (bs, 11H), 3.37 (d, J=8.1 Hz, 1H), 3.23 (d,J=11.1 Hz, 1H), 3.02 (dd, J₁=7.8 Hz, J₂=18 Hz, 1H), 2.69–2.59 (m, 4H),2.35 (s, 3H), 2.26 (s, 6H), 2.00 (s, 3H), 1.76–0.72 (m, 30H).

¹³C NMR (75 MHz, CDCl₃) δ 173.1, 171.5, 168.2, 147.9, 144.7, 142.5,140.7, 140.3, 130.9, 130.6, 127.7, 123.3, 120.0, 117.5, 113.1, 111.9,101.6, 60.5, 59.0, 57.3, 56.7, 55.2, 55.0, 41.6, 39.9, 37.2, 33.5, 33.0,32.9, 32.9, 32.8, 32.5, 32.4, 31.9, 31.7, 29.7, 29.3, 26.6, 26.5, 26.2,24.9, 20.3, 15.8, 14.1, 9.4.

ESI-MS m/z: Calcd. for C₄₆H₆₀N₄O₈: 796.4. Found (M+1)⁺: 797.5.

Example 148

To a solution of 72 (111 mg, 0.162 mmol) in CH₂Cl₂ (0.81 mL), DIPEA(56.3 mL, 0.324 mmol), butyryl chloride (33.6 mL, 0.324 mmol) and DMAP(1.96 mg, 0.016 mmol) were added at 0° C. and the reaction mixture wasstirred for 5 h at this temperature. Then, the solution was diluted withCH₂Cl₂ (10 mL) and washed successively with 0.1 N HCl (5 mL) and asolution of 10% NaHCO₃ (5 ml). The organic layer was dried over Na₂SO₄,filtered, and the solvent was eliminated under reduced pressure. Theresidue was purified by flash column chromatography (RP-18, CH₃CN: H₂O1: 1) to afford 186 (65.4 mg, 54%) as a white solid.

Rf=0.21 Hex:EtOAc 1:2.

¹H NMR (300 MHz, CDCl₃) δ 7.24–7.15 (m, 3H), 7.12–7.04 (m, 2H), 6.84 (s,1H), 5.98 (d, J=1.2 Hz, 1H), 5.92 (d, J=1.2 Hz, 1H), 4.97 (t, J=5.7 Hz,1H), 4.03 (m, 3H), 3.63 (d, J=2.7 Hz, 1H), 3.50 (m, 2H), 3.44 (s, 3H),3.37 (d, J=8.4 Hz, 1H), 3.24 (dt, J₁=2.7 Hz, J₂=11.7 Hz, 1H), 3.02 (dd,J₁=8.1 Hz, J₂=18.3 Hz, 1H), 2.65–2.54 (m, 7H), 2.35 (s, 3H), 2.25 (s,3H), 2.07 (s, 3H), 2.02 (s, 3H), 1.87–1.75 (m, 3H), 1.08 (t, J=7.5 Hz,3H).

¹³C NMR (75 MHz, CDCl₃) δ 171.7, 170.8, 168.2, 147.8, 144.7, 142.5,140.8, 140.6, 140.3, 131.1, 130.5, 128.3, 128.2, 127.6, 126.0, 123.2,117.5, 112.9, 111.8, 101.6, 60.2, 59.0, 57.3, 56.6, 55.1, 54.9, 41.5,39.9, 37.8, 36.0, 31.0, 26.5, 24.8, 22.6, 20.2, 18.5, 15.6, 13.7, 9.3.

ESI-MS m/z: Calcd. for C₄₁H₄₆N₄O₈: 722.83. Found (M+1)⁺: 723.2.

Example 149

To a solution of 72 (80 mg, 0.122 mmol) in CH₂Cl₂ (0.61 mL), DIPEA (64.0mL, 0.367 mmol), hexanoyl chloride (49.5 mL, 0.367 mmol) and DMAP (1.50mg, 0.012 mmol) were added at 0° C. and the reaction mixture was stirredat this temperature for 5 h. Then, the solution was diluted with CH₂Cl₂(10 mL) and washed successively with 0.1 N HCl (5 mL) and a solution of10% NaHCO₃ (5 ml). The organic layer was dried over Na₂SO₄, filtered,and the solvent was eliminated under reduced pressure. The residue waspurified by flash column chromatography (RP-18, CH₃CN: H₂O 6:4) toafford 187 (86.1 mg, 94%) as a white solid.

Rf=0.25 Hex:EtOAc 1:2

¹H NMR (300 MHz, CDCl₃) δ 7.20–7.06 (m, 3H), 6.99–6.97 (m, 2H), 6.77 (s,1H), 5.91 (s, 1H), 5.85 (s, 1H), 4.90 (m, 1H), 3.96 (d, J=3 Hz, 2H),3.57–3.55 (m, 1H), 3.43 (bs, 2H), 3.36 (bs, 3H), 3.29 (brd, J=10.5 Hz,1H), 3.18 (d, J=11.7 Hz, 1H), 2.97 (dd, J₁=4.8 Hz, J₂=12 Hz, 1H),2.58–2.46 (m, 6H), 2.28 (s, 3H), 2.18 (s, 3H), 2.00 (s, 3H), 1.95 (s,3H), 1.86–1.66 (m, 7H), 1.41–1.38 (m, 2H), 0.86–0.81 (m, 3H).

¹³C NMR(75 MHz, CDCl₃) δ 171.7, 171.0, 168.2, 147.8, 144.7, 142.5,140.8, 140.6, 140.3, 131.1, 130.5, 128.3, 128.2, 127.6, 126.0, 117.5,112.9, 111.8, 101.6, 60.2, 59.0, 57.3, 56.6, 55.1, 55.0, 41.5, 39.9,37.8, 34.1, 31.3, 31.1, 29.6, 24.8, 24.7, 22.3, 20.2, 15.6, 13.8.

ESI-MS m/z: Calcd. for C₄₃H₅₀N₄O₈: 750.88. Found (M+1)⁺: 751.3.

Example 150

To a solution of 85 (80 mg, 0.110 mmol) in CH₂Cl₂ (0.55 mL), DIPEA (57.7mL, 0.331 mmol), butyryl chloride (34.4 mL, 0.331 mmol) and DMAP (1.30mg, 0.011 mmol) were added at 0° C. and the reaction mixture was stirredat 23° C. for 5 h. Then, the solution was diluted with CH₂Cl₂ (10 mL)and washed successively with 0.1 N HCl (5 mL) and a solution of 10%NaHCO₃ (5 ml). The organic layer was dried over Na₂SO₄, filtered, andthe solvent was eliminated under reduced pressure. The residue waspurified by flash column chromatography (RP-18, CH₃CN: H₂O 1:1) toafford 188 (70.1 mg, 80%) as a white solid.

Rf=0.54 MeOH:EtOAc 1:5.

¹H NMR (300 MHz, CDCl₃) δ 7.28–7.14 (m, 5H), 6.80 (s, 1H), 6.07 (d,J=6.6 Hz, 1H), 6.00 (d, J=1.5 Hz, 1H), 5.90 (d, J=1.5 Hz, 1H), 5.35 (t,J=5.4 Hz, 1H), 4.12 (d, J=2.4 Hz, 1H), 4.05 (bs, 1H), 3.89 (brt, J=6.9Hz, 1H), 3.66 (s, 3H), 3.64–3.63 (m, 1H), 3.59–3.45 (m, 2H), 3.40 (brd,J=7.8 Hz, 1H), 3.20 (dt, J₁=2.7 Hz, J₂=12 Hz, 1H), 3.00 (dd, J₁=8.1 Hz,J₂=18 Hz, 1H), 2.87 (t, J=8.1 Hz, 2H), 2.71 (d, J=18.6 Hz, 1H),2.66–2.61 (m, 1H), 2.58 (t, J=7.2 Hz, 2H), 2.41–2.35 (m, 2H), 2.33 (s,3H), 2.23 (s, 3H), 2.21 (s, 3H), 2.00 (s, 3H), 1.90–1.77 (m, 3H), 1.08(t, J=7.2 Hz, 3H), 0.69 (d, J=6.9 Hz, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 172.0, 171.3, 170.8, 168.5, 147.7, 144.7,142.5, 140.6, 140.5, 140.3, 131.0, 130.7, 128.4, 128.2, 127.7, 126.1,123.1, 120.3, 117.5, 112.7, 111.8, 101.6, 60.3, 59.1, 57.3, 57.2, 55.4,54.9, 48.2, 41.5, 39.5, 38.0, 36.0, 31.4, 26.8, 26.6, 24.6, 20.1, 18.5,18.1, 15.7, 13.7, 9.2.

ESI-MS m/z: Calcd. for C₄₄H₅N₅O₉: 793.9. Found (M+1)⁺: 794.3.

Example 151

To a solution of 85 (80 mg, 0.1101 mmol) in CH₂Cl₂ (0.55 mL), DIPEA(57.7 mL, 0.331 mmol), hexanoyl chloride (46.3 mL, 0.331 mmol) and DMAP(1.30 mg, 0.011 mmol) were added at 0° C. and the reaction mixture wasstirred at 23° C. for 5 h. Then the solution was diluted with CH₂Cl₂ (10mL) and washed succesively with 0.1 N HCl (5 mL) and a solution of 10%NaHCO₃ (5 ml). The organic layer was dried over Na₂SO₄, filtered, andthe solvent was eliminated under reduced pressure. The residue waspurified by column chromatography (RP-18, CH₃CN: H₂O 1:1) to afford 189(80 mg, 88%) as a white solid.

Rf=0.23 Hex:EtOAc 1:3.

¹H NMR (300 MHz, CDCl₃) δ 7.21–7.08 (m, 5H), 6.74 (s, 1H), 6.00 (d,J=6.9 Hz, 1H), 5.94 (d, J=1.5 Hz, 1H), 5.84 (d, J=1.5 Hz, 1H), 5.24 (t,J=5.4 Hz, 1H), 4.06 (bs, 1H), 4.00 (bs, 1H), 3.83 (t, J=6 Hz, 1H), 3.59(s, 3H), 3.57 (m, 1H), 3.53–3.40 (m, 2H), 3.33 (d, J=7.8 Hz, 1H), 3.14(d, J=11.7 Hz, 1H), 2.94 (dd, J₁=8.4 Hz, J₂=18 Hz, 1H), 2.81 (t, J=7.5Hz, 2H), 2.65 (d, J=18 Hz, 1H), 2.60–2.54 (m, 1H), 2.52 (t, J=7.2 Hz,2H), 2.35–2.29 (m, 2H), 2.27 (s, 3H), 2.17 (s, 3H), 2.15 (s, 3H), 1.95(s, 3H), 1.76–1.60 (m, 3H), 1.35–1.29 (m, 2H), 1.84 (m, 2H), 0.85–0.78(m, 3H), 0.62 (t, J=6.6 Hz, 3H).

¹³C NMR(75 MHz, CDCl₃) δ 172.0, 171.3, 171.1, 168.4, 147.8, 144.8,142.6, 140.7, 140.5, 131.2, 130.6, 128.4, 128.3, 127.7, 126.2, 123.1,120.3, 117.5, 112.6, 112.0, 101.7, 60.4, 59.1, 57.4, 57.2, 55.4, 54.9,48.3, 41.5, 39.6, 38.1, 34.1, 33.6, 31.5, 31.3, 26.7, 24.7, 22.3, 20.2,18.2, 15.7, 13.9, 9.3.

ESI-MS m/z: Calcd. for C₄₆H₅₅N₅O₉: 821.96. Found (M+1)⁺: 822.3.

Example 152

To a solution of 53 (100 mg, 0.145 mmol) in CH₂Cl₂ (0.72 mL), DIPEA(50.6 mL, 0.291 mmol) and acetyl chloride (20.7 mL, 0.291 mmol) wereadded at 0° C. and the reaction mixture was stirred for 4 h at 23° C.Then, the solution was diluted with CH₂Cl₂ (10 mL), and washedsuccessively with 0.1 N HCl (5 mL), and a solution of 10% NaHCO₃ (5 ml).The organic layer was dried over Na₂SO₄, filtered, and the solvent waseliminated under reduced pressure. The residue was purified by flashcolumn chromatography (SiO₂, Hex: EtOAc 1:2) to afford 190 (27 mg, 25%)as a white solid.

Rf=0.24 Hex:EtOAc 1:1.

¹H NMR (300 MHz, CDCl₃) δ 6.82 (s, 1H), 6.02 (d, J=0.9 Hz, 1H), 5.92 (d,J=0.9 Hz, 1H), 5.30 (bs, 1H), 4.14 (d, J=2.7 Hz, 1H), 4.10 (s, 1H),3.90–3.73 (m, 2H), 3.68 (s, 3H), 3.67 (bs, 1H), 3.49 (bs, 1H), 3.42(brd, J=8.1 Hz, 1H), 3.24–3.20 (m, 1H), 3.01 (dd, J₁=8.4 Hz, J₂=18.3 Hz,1H), 2.78 (d, J=18 Hz, 1H), 2.64 (brd, J=15.6 Hz, 1H), 2.36 (s, 3H),2.34 (s, 3H), 2.24 (s, 3H), 2.20 (s, 3H), 2.02 (s, 3H), 1.77 (dd,J₁=11.7 Hz, J₂=15.6 Hz, 1H), 0.65 (d, J=6.6 Hz, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 170.2, 168.6, 168.1, 167.6, 147.9, 144.9,142.8, 140.5, 131.5, 131.0, 127.7, 123.2, 120.3, 117.5, 112.3, 112.2,101.7, 60.4, 59.0, 57.4, 57.2, 55.2, 54.9, 48.6, 41.5, 39.1, 36.6, 29.7,26.7, 24.6, 20.7, 20.2, 17.6, 15.5, 9.2.

ESI-MS m/z: Calcd. for C₃₅H₃₈F₃N₅O₉: 729.70. Found (M+1)⁺: 730.3.

Example 153

To a solution of 53 (150 mg, 0.218 mmol) in CH₂Cl₂ (1.09 mL). DIPEA(151.9 mL, 0.87 mmol), butyryl chloride (90.6 mL, 0.87 mmol) and DMAP(2.70 mg, 0.02 mmol) were added at 0° C. and the reaction mixture wasstirred at 23° C. for 4 h. Then, the solution was diluted with CH₂Cl₂(10 mL) and washed successively with 0.1 N HCl (5 mL) and a solution of10% NaHCO₃ (5 ml). The organic layer was dried over Na₂SO₄, filtered,and the solvent was eliminated under reduced pressure. The residue waspurified by flash column chromatography (RP-18, CH₃CN: H₂O 4:1) toafford 191 (20.2 mg, 12%) as a white solid.

Rf=0.3 Hex:EtOAc 1:1.

¹H NMR (300 MHz, CDCl₃) δ 6.81 (s, 1H), 6.03 (d, J=1.2 Hz, 1H), 5.92 (d,J=1.2 Hz, 1H), 5.16 (t, J=5.4 Hz, 1H), 4.13 (d, J=2.1 Hz, 1H), 4.10 (bs,1H), 3.87–3.82 (m, 1H), 3.80–3.74 (m, 1), 3.68 (s, 3H), 3.64 (d, J=3 Hz,1H), 3.52–3.47 (m, 1H), 3.42 (brd, J=7.2 Hz, 1H), 3.24–3.20 (m, 1H),3.02 (dd, J₁=8.1 Hz, J₂=18.3 Hz, 1H), 2.77 (d, J=17.7 Hz, 1H), 2.64(brd, J=16.2 Hz, 1H), 2.58 (t, J=7.2 Hz, 2H), 2.33 (s, 3H), 2.25 (s,3H), 2.22 (s, 3H), 2.02 (s, 3H), 1.87–1.73 (m, 3H), 1.08 (t, J=7.2 Hz,3H), 0.68 (d, J=6.6 Hz, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 172.8, 172.1, 170.4, 157.8, 150.0, 146.9,144.8, 142.6, 142.5, 133.3, 132.8, 129.6, 125.3, 122.3, 119.5, 118.4,115.7, 114.3, 114.2, 103.8, 62.4, 61.0, 59.4, 59.2, 57.2, 57.0, 50.6,43.6, 41.2, 38.1, 31.7, 28.7, 26.6, 22.2, 20.6, 19.7, 17.5, 15.7, 11.2.

ESI-MS m/z: Calcd. for C₃₇H₂F₃N₅O₉: 757.75. Found: 758.5 (M+1)⁺, 780.5(M+23)⁺.

Example 154

To a solution of 53 (150 mg, 0.218 mmol) in CH₂Cl₂ (1.09 mL), DIPEA(151.9 mL, 0.87 mmol), acetyl chloride (62.0 mL, 0.87 mmol) and DMAP(2.70 mg, 0.02 mmol) were added at 0° C. and the reaction mixture wasstirred at 23° C. for 5 h. Then, the solution was diluted with CH₂Cl₂(10 mL) and washed successively with 0.1 N HCl (5 mL) and a solution of10% NaHCO₃ (5 ml). The organic layer was dried over Na₂SO₄, filtered,and the solvent was eliminated under reduced pressure. The residue waspurified by flash column chromatography (RP-18, CH₃CN: H₂O 1:1) toafford 192 (111 mg, 62%) as a white solid.

Rf=0.25 Hex:EtOAc 1:1.

¹H NMR (300 MHz, CDCl₃) δ 6.80 (s, 1H), 5.87 (s, 1H), 5.81 (s, 1H), 4.70(dd, J₁=2.4 Hz, J₂=9.9 Hz, 1H), 4.20 (d, J=6.3 Hz, 1H), 4.09 (s, 1H),3.74 (s, 31H), 3.60 (s, 1H), 3.28 (d, J=7.5 Hz, 1H), 3.17 (d, J=12 Hz,1H), 3.07 (dd, J₁=7.2 Hz, J₂=18.3 Hz, 1H), 2.93 (d, J=13.2 Hz, 1H), 2.66(d, J=15.3 Hz, 1H), 2.53 (d, J=17.7 Hz, 1H), 2.47–2.20 (m, 1H), 2.37 (s,1H), 2.33 (s, 3H), 2.26 (s, 3H), 2.24 (s, 3H), 2.08 (s, 3H), 2.00 (s,3H), 1.96 (s, 3H), 1.72 (t, J=14.4 Hz, 1H), 1.53 (d, J=6.9 Hz, 3H).

¹³C NMR(75 MHz, CDCl₃) δ 174.1, 168.6, 168.4, 167.5, 147.7, 144.8,142.2, 140.4, 131.1, 130.5, 126.9, 123.3, 120.4, 117.5, 112.4, 111.8,101.1, 60.7, 60.6, 57.6, 57.2, 56.6, 55.3, 52.7, 48.3, 41.5, 31.6, 29.7,26.4, 25.5, 23.0, 22.6, 20.7, 20.5, 20.2, 17.8, 15.9, 14.1, 9.5.

ESI-MS m/z: Calcd. for C₃₉H₄₂F₃N₅O₁₁: 813.7. Found (M+1)⁺: 814.3.

Example 155

To a solution of 53 (150 mg, 0.218 mmol) in CH₂Cl₂ (1.09 mL), DIPEA(151.9 mL, 0.87 mmol), butyryl chloride (90.6 mL, 0.87 mmol) and DMAP(2.70 mg, 0.02 mmol) were added at 0° C. and the reaction mixture wasstirred at 23° C. for 4 h. Then, the solution was diluted with CH₂Cl₂(10 mL) and washed successively with 0.1 N HCl (5 mL) and a solution of10% NaHCO₃ (5 ml). The organic layer was dried over Na₂SO₄, filtered,and the solvent was eliminated under reduced pressure. The residue waspurified by flash column chromatography (RP-18, CH₃CN: H₂O 4:1) toafford 193 (58 mg, 30%) as a white solid.

Rf=0.38 Hex:EtOAc 1:1.

¹H NMR (300 MHz, CDCl₃) δ 6.85 (s, 1H), 5.99 (d, J=1.2 Hz, 1H), 5.90 (d,J=1.2 Hz, 1H), 5.47–5.42 (m, 2H), 4.09–4.08 (m, 2H), 3.69 (s, 3H), 3.66(m, 1H), 3.41 (d, J=7.5 Hz, 1H), 3.28–3.18 (m, 2H), 3.07 (dd, J₁=8.1 Hz,J₂=18 Hz, 1H), 2.66 (d, J=18.6 Hz, 1H), 2.61–2.39 (m, 3H), 2.34 (s, 3H),2.26 (s, 3H), 2.21 (s, 3H), 2.01 (s, 3H), 1.95–1.79 (m, 6H), 1.72–1.59(m, 6H) 1.09 (t, J=7.5 Hz, 3H), 0.99–0.94 (m, 6H), 0.85 (d, J=6.9 Hz,3H).

¹³C NMR(75 MHz, CDCl₃) δ 171.2, 170.7, 169.1, 168.4, 148.1, 145.0,142.7, 140.9, 140.6, 131.2, 130.5, 128.4, 123.4, 119.9, 117.6, 113.0,112.1, 101.9, 60.7, 59.5, 57.6, 56.5, 55.7, 55.2, 41.8, 41.4, 36.3,35.8, 29.9, 27.0, 25.3, 20.5, 20.0, 18.8, 18.3, 15.8, 14.0, 13.8, 13.4,12.7, 9.6.

ESI-MS m/z: Calcd. for C₄₅H₅₄F₃N₅O₁₁: 897.93. Found (M+1)⁺: 898.3.

Example 156

To a solution of 53 (150 mg, 0.218 mmol) in CH₂Cl₂ (1.09 mL), DIPEA(151.9 mL, 0.87 mmol), hexanoyl chloride (121.9 mL, 0.87 mmol) and DMAP(2.70 mg, 0.02 mmol) were added at 0° C. and the reaction mixture wasstirred at 23° C. for 4 h. Then, the solution was diluted with CH₂Cl₂(10 mL) and washed successively with 0.1 N HCl (5 mL) and a solution of10% NaHCO₃ (5 ml). The organic layer was dried over Na₂SO₄, filtered,and the solvent was eliminated under reduced pressure. The residue waspurified by flash column chromatography (RP-18, CH₃CN: H₂O 4:1) toafford 194 (37.5 mg, 22%) as a white solid.

Rf=0.32 Hex:EtOAc 1:1.

¹H NMR (300 MHz, CDCl₃) δ 6.80 (s, 1H), 6.02 (d, J=1.2 Hz, 1H), 5.92 (d,J=1.2 Hz, 1H), 5.22 (t, J=5.7 Hz, 1H), 4.13 (d, J=2.4 Hz, 1H), 4.09 (s,1H), 3.88–3.81 (m, 1H), 3.80–3.71 (m, 1H), 3.67 (s, 3H), 3.64 (d, J=3Hz, 1H), 3.52–3.43 (m, 1H), 3.41 (brd, J=6.6 Hz, 1H), 3.23–3.19 (m, 1H),3.00 (dd, J₁=8.7 Hz, J₂=18.6 Hz, 1H), 2.77 (d, J=18 Hz, 1H), 2.67–2.56(m, 3H), 2.33 (s, 3H), 2.24 (s, 3H), 2.22 (s, 3H), 2.01 (s, 3H),1.82–1.74 (m, 4H), 1.43–1.38 (m, 3H), 0.97–0.88 (m, 3H), 0.67 (d, J=6.9Hz, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 171.2, 170.3, 168.6, 148.2, 145.1, 143.0,140.8, 140.7, 131.7, 131.1, 127.8, 123.5, 120.6, 117.7, 112.5, 102.0,60.7, 59.2, 57.6, 57.4, 55.4, 55.2, 48.9, 41.8, 34.4, 31.8, 31.6, 29.9,26.9, 25.0, 24.8, 22.9, 22.5, 20.4, 17.9, 15.8, 14.3, 14.1, 9.5.

ESI-MS m/z: Calcd. for C₃₉H₄₆F₃N₅O₉: 785.81. Found: 786 (M+1)⁺, 805.5(M+23)⁺.

Example 157

To a solution of 53 (150 mg, 0.218 mmol) in CH₂Cl₂ (1.09 mL), DIPEA(75.9 mL, 0.436 mmol), and decanoyl chloride (92.7 mL, 0.436 mmol) wereadded at 0° C. and the reaction mixture was stirred at 23° C. for 4 h.Then, the solution was diluted with CH₂Cl₂ (10 mL) and washedsuccessively with 0.1 N HCl (5 mL), and a solution of 10% NaHCO₃ (5 ml).The organic layer was dried over Na₂SO₄, filtered, and the solvent waseliminated under reduced pressure. The residue was purified by flashcolumn chromatography (RP-18, CH₃CN: H₂O 1:1) to afford 195 (75 mg, 41%)as a white solid.

Rf=0.32 Hex:EtOAc 1:1.

¹H NMR (300 Hz, CDCl₃) δ 6.82 (s, 1H), 6.03 (d, J=1.5 Hz, 1H), 5.93 (d,J=1.5 Hz, 1H), 5.26 (bs, 1H), 4.15 (s, 1H), 4.11 (s, 1H), 3.89–3.75 (m,2H), 3.68 (s, 3H), 3.65 (bs, 1H), 3.52–3.44 (m, 1H), 3.43 (d, J=8.1 Hz,1H), 3.22 (brd, J=11.4 Hz, 1H), 3.03 (dd, J₁=7.8 Hz, J₂=17.4 Hz, 1H),2.78 (d, J=17.7 Hz, 1H), 2.69–2.56 (m, 3H), 2.34 (s, 3H), 2.26 (s, 3H),2.23 (s, 3H), 2.03 (s, 3H), 1.83–1.74 (m, 3H), 1.83–1.74 (m, 12H),0.90–8.88 (m, 3H), 0.68 (d, J=6 Hz, 3H).

¹³C NMR(75 Hz, CDCl₃) δ 171.0, 170.1, 168.4, 148.0, 144.8, 142.8, 140.5,131.5, 130.8, 127.5, 123.3, 120.3, 117.5, 112.3, 112.2, 101.7, 60.4,59.0, 57.4, 57.2, 55.1, 55.0, 48.6, 41.5, 39.1, 34.2, 31.8, 29.4, 29.2,26.7, 25.0, 24.6, 22.6, 20.2, 17.6, 15.5, 14.0, 9.2.

ESI-MS m/z: Calcd. for C₄₃H₅₄F₃N₅O₉: 841.91. Found (M+1)⁺: 842.3.

Example 158

To a solution of 53 (150 mg, 0.218 mmol) in CH₂Cl₂ (1.09 mL), DIPEA(75.9 mL, 0.436 mmol), and stearoyl chloride (147.3 mL, 0.436 mmol) wereadded at 0° C. and the reaction mixture was stirred at 23° C. for 4 h.Then, the solution was diluted with CH₂Cl₂ (10 mL) and washedsuccessively with 0.1 N HCl (5 mL) and a solution of 10% NaHCO₃ (5 ml).The organic layer was dried over Na₂SO₄, filtered, and the solvent waseliminated under reduced pressure. The residue was purified by flashcolumn chromatography (RP-18, CH₃CN: H₂O 1:1) to afford 196 (86 mg, 41%)as a white solid.

Rf=0.42 Hex:EtOAc 1:1.

¹H NMR (300 MHz, CDCl₃) δ 6.81 (s, 1H), 6.03 (s, 1H), 5.92 (s, 1H), 5.21(bs, 1H), 4.14 (s, 1H), 4.10 (s, 1H), 3.88–3.74 (m, 2H), 3.67 (s, 3H),3.64 (d, J=3 Hz, 1H), 3.49 (brd, J=14.7 Hz, 1H), 3.42 (d, J=8.1 Hz, 1H),3.22 (brd, J=11.4 Hz, 1H), 3.02 (dd, J₁=8.7 Hz, J₂=18.6 Hz, 1H), 2.78(d, J=18 Hz, 1H), 2.68–2.56 (m, 3H), 2.33 (s, 3H), 2.25 (s, 3H), 2.02(s, 3H), 1.82–1.73 (m, 3H), 1.42–1.19 (m, 28H), 0.87 (t, J=7.2 Hz, 3H),0.67 (d, J=6.6 Hz, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 171.0, 170.2, 168.5, 147.9, 144.8, 142.8,140.4, 131.4, 130.9, 127.5, 123.3, 120.4, 117.5, 112.4, 112.1, 101.7,60.4, 58.9, 57.4, 57.2, 55.2, 55.0, 48.6, 41.5, 39.0, 34.2, 31.9, 29.7,29.6, 29.4, 29.3, 29.2, 26.7, 25.1, 24.6, 22.7, 20.2, 17.6, 15.5, 14.1,9.2. ESI-MS m/z: Calcd. for C₅₁H₇₀F₃N₅O₉: 953.5. Found (M+1)⁺: 954.4.

Example 159

To a solution of 45 (10 mg, 0.019 mmol) in CH₂Cl₂ (0.095 mL),triethylamine (2.94 mL, 0.021 mmol) and allyl bromide (2.0 mL, 0.023mmol) were added at 23° C. The reaction mixture was stirred for 6 h andthen, the solvent was removed under reduced pressure. The residue waspurified by flash column chromatography (SiO₂, MeOH: EtOAc 1:5) toafford 197 (3.8 mg, 35%) as a white solid.

Rf=0.19 EtOAc:MeOH 5:1.

¹H NMR (300 MHz, CDCl₃) δ 6.43 (s, 1H), 5.95 (s, 1H), 5.89 (s, 1H),5.62–5.59 (m, 1H), 4.94–4.84 (m, 2H), 4.19 (s, 1H), 4.08 (s, 1H), 3.98(t, J=4.5 Hz, 1H), 3.76 (s, 3H), 3.32–3.26 (m, 2H), 3.07 (dd, J₁=7.5 Hz,J₂=17.4 Hz, 1H), 2.89 (d, J=6 Hz, 2H), 2.80 (d, J=3.9 Hz, 1H), 2.76 (d,J=3.3 Hz, 1H), 2.57–2.52 (m, 2H), 2.33 (s, 6H), 2.24 (s, 3H), 1.99 (s,3H), 1.88–1.79 (dd, J₁=12.9 Hz, J₂=15.9 Hz, 1H).

ESI-MS m/z: Calcd. for C₃₁H₃₆N₄O₆: 560.64. Found (M+1)⁺: 561.3.

Example 160

To a solution of 146 (50 mg, 0.096 mmol) in CH₂Cl₂ (0.96 mL), pyridine(11.7 mL, 0.144 mmol), and cinnamoyl chloride (24.0 mg, 0.144 mmol) wereadded at 23° C. and the reaction mixture was stirred for 18 h at thattemperature. Then, the solution was diluted with CH₂Cl₂ (10 mL) andwashed successively with 0.1 N HCl (5 mL) and a solution of 10% NaHCO₃(5 ml). The organic layer was dried over Na₂SO₄, filtered, and thesolvent was eliminated under reduced pressure. The residue was purifiedby flash column chromatography (SiO₂, Hex:EtOAc 1:2) to afford 198 (54mg, 86%) as a white solid.

Rf=0.45 Hex:EtOAc 1:1.

¹H NMR (300 MHz, CDCl₃) δ 7.41–7.37 (m, 6H), 6.38 (s, 1H), 6.19–6.03 (m,1H), 6.08 (d, J=15.9 Hz, 1H), 5.93 (d, J=1.5 Hz, 1H), 5.88 (d, J=1.5 Hz,1H), 5.62 (s, 1H), 5.38 (dd, J₁=1.5 Hz, J₂=17.1 Hz, 1H), 5.26 (dd,J₁=1.5 Hz, J₂=10.5 Hz, 1H), 4.47 (dd, J₁=3.6 Hz, J₂=10.8 Hz, 1H),4.23–4.11 (m, 5H), 3.89 (dd, J₁=4.8 Hz, J₂=11.1 Hz, 1H), 3.51 (s, 3H),3.34 (brd, J=8.4 Hz, 1H), 3.27–3.21 (m, 2H), 2.97 (dd, J₁=7.8 Hz,J₂=17.7 Hz, 1H), 2.28 (s, 3H), 2.15 (s, 3H), 2.04 (s, 3H), 1.91 (dd,J₁=12 Hz, J₂=15.6 Hz, 1H).

¹³C NMR (75 MHz, CDCl₃) δ 166.5, 148.8, 146.7, 144.7, 144.5, 142.7,139.5, 134.4, 134.1, 131.1, 130.6, 129.1, 128.7, 128.2, 121.9, 121.2,118.5, 117.8, 116.8, 112.9, 112.7, 101.5, 74.7, 65.2, 60.7, 60.6, 57.4,56.8, 56.6, 55.7, 41.9, 31.8, 26.7, 25.5, 22.9, 15.9, 14.4, 9.7.

ESI-MS m/z: Calcd. for C₃₈H₃₉N₃O₇: 649.7. Found (M+1)⁺: 650.3.

Example 161

To a solution of 161 (78.5 mg, 0.146 mmol) and the cysteine derivative(81.1 mg, 0.247 mmol) in anhydrous CH2Cl2 (7.3 mL), DMAP (50 mg, 0.41mmol) and EDC.HCl (78.1 mg, 0.41 mmol) were added at 23° C. The reactionmixture was stirred at 23° C. under Argon atmosphere for 1.5 h. Themixture was diluted with CH2Cl2 (20 mL) and extracted with an aqueoussaturated solution of sodium bicarbonate (25 mL). The aqueous phase wasextracted with additional CH2Cl2 (20 mL) and the combined organicextracts were dried over Na2SO4, filtered and the solvent was eliminatedunder reduced pressure. The crude of the reaction was purified by flashcolumn chromatography (inner diameter of the column 2 cm, height ofsilica 10 cm) with mixtures of ethyl acetate/hexane in a gradientmanner, from 1:4 to 3:1 as eluent. Compound 199 (113 mg, 88%) wasobtained as a pale yellow solid.

Rf=0.36 Hex:EtOAc 1:1.

¹H NMR (300 MHz, CDCl₃) δ: 7.76 (d, J=7.8 Hz, 2H), 7.63 (d, J=7.8 Hz,2H), 7.40 (t, J=7.6 Hz, 2H), 7.29 (t, J=7.6 Hz, 2H), 6.54 (s, 1H), 5.80(s, 1H), 5.74 (s, 1H), 5.10 (d, J=5.7 Hz, 1H), 5.08 (d, J=5.7 Hz, 1H),4.50 (dd, J=4.9 Hz, J=11.8 Hz, 1H), 4.20–4.05 (m, 4H), 4.02 (s, 3H),3.81 (s, 3H), 3.61 (d, J=13.8 Hz, 1H), 3.55 (d, J=13.8 Hz, 1H), 3.50 (s,3H), 3.21 (m, 1H), 3.06 (m, 1H), 3.00 (d, J=6.0 Hz, 2H), 2.90 (dd, J=8.9Hz, J=17.4 Hz, 1H), 2.79 (s, 1H), 2.56 (m, 1H), 2.50 (dd, J=4.8 Hz,J=14.9 Hz, 1H), 2.21 (s, 3H), 2.18 (s, 3H), 1.80 (s, 3H), 1.75 (m, 2H).

ESI-MS m/z: Calcd. for C₄₆H₈N₄O₁₀S: 848.3. Found: 849.3 (M+1)⁺, 871.3(M+23)⁺. HPLC: Conditions: Column: Simmetry C18, Mobile phase: CH₃CN/H₂Oin gradient from 50 to 100% in 25 minutes. ø=1 mL/min, t=40° C.Retention time: 16.04 minutes. HPLC purity in area: 89.29%.

Example 162

To a solution of 161 (80 mg, 0.148 mmol) and the cysteine derivative (76mg, 0.223 mmol) in anhydrous CH₂Cl₂ (6.8 mL), DMAP (45 mg, 0.37 mmol)and EDC.HCl (71 mg, 0.37 mmol) were added at 23° C. The reaction mixturewas stirred at 23° C. under Argon atmosphere for 2.5 h Then, the mixturewas diluted with CH₂Cl₂ (20 mL) and extracted with an aqueous saturatedsolution of sodium bicarbonate (25 mL). The aqueous phase was extractedwith additional CH₂Cl₂ (20 mL) and the combined organic extracts weredried over Na₂SO₄, filtered and the solvent was eliminated under reducedpressure. The crude of the reaction was purified by flash columnchromatography (inner diameter of the column 2 cm, height of silica 10cm) with mixtures of ethyl acetate/hexane in gradient from 1:4 to 3:1 aseluent. Compound 200 (83 mg, 65%) was obtained as a pale yellow solid.

Rf=0.5 Hex:EtOAc 1:1.

¹H NMR (300 MHz, CDCl₃) δ: 7.71 (m, 3H), 7.49 (d, J=7.3 Hz, 1H), 7.36(t, J=7.3 Hz, 2H), 7.32–7.23 (m, 2H), 6.65 (s, 1H), 5.80 (s, 1H), 5.79(s, 1H), 5.13 (d, J=6.1 Hz, 1H), 5.11 (d, J=6.1 Hz, 1H), 5.05 (d, J=6.1Hz, 1H), 5.01 (d, J=6.3 Hz, 1H), 4.76 (dd, J=3.9 Hz, J=11.9 Hz, 1H),4.15–4.03 (m, 4H), 3.96 (t, J=4.0 Hz, 1H), 3.87 (s, 3H), 3.55 (s, 3H),3.51 (s, 3H), 3.34–3.29 (m, 2H), 3.24 (dd, J=5.5 Hz, J=13.5 Hz, 1H),3.03 (m, 1H), 2.97 (t, J=7.5 Hz, 1H), 2.44–2.35 (m, 3H), 2.29 (s, 3H),2.14 (s, 3H), 1.98 (dd, J=8.06, J=15.1 Hz, 2H), 1.75 (s, 3H).

¹³C NMR (75 MHz, CDCl₃) δ 196.98, 161.13, 158.21, 149.01, 148.78,145.05, 144.91, 141.01, 140.69, 140.07, 137.53, 132.76, 131.15, 129.41,127.70, 127.67, 127.21, 126.83, 125.28, 125.05, 124.94, 122.51, 119.84,119.73, 116.61, 110.26, 104, 57, 101.40, 99.23, 96.70, 70.25, 63.15,60.40, 58.89, 57.52, 56.98, 56.72, 56.15, 55.06, 47.22, 41.37, 38.26.35.22, 29.57, 25.34, 15.62, 7.26.

ESI-MS m/z: Calcd. for C₄₇H₄₉N₃O₁₁S: 863.97. Found: 865.0 (M+1)⁺, 887.1(M+23)⁺. HPLC: Conditions: Column: Simmetry C18. Mobile phase: CH₃CN/H₂Oin gradient from 50 to 100% in 25 minutes. ø=1 mL/min. t=40° C.Retention time: 15.36 minutes. HPLC purity in area: 91.56%.

Example 163

To a solution of 161 (418 mg, 0.77 mmol) and the cysteine derivative(321 mg, 0.77 mmol) in anhydrous CH₂Cl₂ (35 mL), DMAP (235 mg, 1.92mmol) and EDC.HCl (369 mg, 1.92 mmol) were added at 23° C. and thereaction was stirred under Argon atmosphere for 2 h. The mixture wasdiluted with CH₂Cl₂ (20 mL) and extracted with an aqueous saturatedsolution of sodium bicarbonate (25 mL). The aqueous phase was extractedwith additional CH₂Cl₂ (20 mL) and the combined organic extracts weredried over Na₂SO₄, filtered and the solvent was eliminated under reducedpressure. The crude of the reaction was purified by flash columnchromatography (inner diameter of the column 3 cm, height of silica 11cm) with mixtures of ethyl acetate/hexane in a gradient manner, from 1:3to 3:1 as eluent. Compound 201 (372 mg, 52%) was obtained as a paleyellow solid.

Rf=0.41 Hex:EtOAc 1:1.

¹H-RMN (CDCl₃, 300 MHz) δ 7.76–7.64 (m, 4H), 7.41–7.30 (m, 4H), 6.54 (s,1H major isomer), 6.51 (s, 1H, minor isomer), 5.69 (s, 1H, minorisomer), 5.67 (s, 1H, major isomer), 5.60 (s, 1H minor isomer), 5.57 (s,1H major isomer), 5.08 (s, 2H), 4.26 (t, J=5.1 Hz, 1H minor isomer),4.23 (t, J=4.9 Hz, 1H major isomer), 4.07–4.03 (m, 3H), 3.98–3.88 (m,3H), 3.84 (s, 3H), 3.71 (dt, J₁=5.6 Hz, J₂=10.0 Hz, 1H), 3.49 (s, 3H,major isomer), 3.49 (s, 3H, minor isomer), 3.40 (dt, J₁=5.6 Hz, J₂=9.5Hz, 1H), 3.18 (m, 3H), 3.11 (m 1H), 2.91–2.82 (m, 1H), 2.48–2.28 (m,2H), 2.24 (s, 3H), 2.16 (s, 3H, major isomer), 2.14 (s, 3H, minorisomer), 2.03 (s, 3H), 1.91 (dt, J₁=8.8 Hz, J₂=14.4 Hz, 1H), 1.76 (s,3H, minor isomer), 1.76 (s, 3H major isomer), 0.85 (s, 9H minor isomer),0.85 (s, 9H major isomer), 0.04 and 0.01 (s, 6H both isomers).

ESI-MS m/z: Calcd. for C₅₁H₆₁N₃O₁₀SSi: 935.4. Found: 936.4 (M+1)⁺, 958.3(M+23)⁺.

Example 164

To a solution of 25 (2 mg, 0.0035 mmol) and an excess amount of thecysteine derivative in anhydrous CH₂Cl₂ (0.2 mL), an excess amounts ofDMAP and EDC.HCl were added at 23° C. The reaction mixture was stirredat 23° C. under Argon atmosphere for 14 h. Then, the mixture was dilutedwith CH₂Cl₂ (10 mL) and washed with a saturated aqueous solution ofsodium bicarbonate (10 mL). The aqueous phase was extracted withadditional CH₂Cl₂ (10 mL). The combined organic layers were dried overNa₂SO₄, filtered and the solvent was eliminated under reduced pressure.The crude of the reaction was purified by flash column chromatography(SiO₂, Hex:EtOAc 4:1) to afford 202 as a pale yellow solid.

¹H NMR (300 MHz, CDCl₃) (poor resolution) δ 7.78.7.62 (m, 4H), 7.41–7.26(m, 4H), 6.73 (s, 1H), 6.10 (m, 1H), 5.92 (d, J=1.3 Hz, 1H), 5.88 (d,J=1.3 Hz, 1H), 5.40–5.22 (m, 2H), 5.11 (s, 3H), 5.02 (d, J=13.8 Hz, 1H),4.29–4.02 (m, 6H), 3.97 (m, 1H), 3.72 (d, J=12.5 Hz, 2H), 3.70 (s, 3H),3.58 (s, 3H), 3.51 (d, J=12.3 Hz, 2H), 3.50 (s, 3H), 3.49–3.20 (m, 4H),2.54–2.28 (m, 4H), 2.40 (s, 3H), 2.21 (s, 3H), 2.16 (s, 3H).

Fermentation Procedures

Example A

Seed medium YMP3 containing 1% glucose; 0.25% beef extract; 0.5%bacto-peptone; 0.25% NaCl; 0.8% CaCO₃ was inoculated with 0.1% of afrozen vegetative stock of the microorganism, strain A2-2 ofPseuidomonas fluorescens, and incubated on a rotary shaker (250 rpm) at27° C. After 30 h of incubation, the seed culture was added to aagitated-vessel fermentor with a production medium composed of 2%dextrose; 4% mannitol, 2% dried brewer's yeast (Vitalevor® Biolux,Belgium); 1% (NH₄)₂SO₄: 0.04% K₂HPO₄; 0.8 KCl; 0.001% FeCl₃; 0.1% L-Tyr;0.8% CO₃Ca; 0.05% PPG-2000; 0.2% anti-foam silicone (ASSAF-100, RHODIAUK). The sterilisation was carried out at 122° C. 30 minutes. The volumeinoculated was a 2% (v/v). The temperature was 27° C. (0 to 16 h) and24° C. from 16 h to final process (41 hours). The dissolveoxygen-pressure was upper to 25%. The pH was controlled at 6.0 withdiluted sulphuric acid since 28 hours till final process. Theoverpressure was 0.5 bar. A 1% mannitol or sorbitol was added from 16 hto final process (for two days running) and 2% for three daysfermentation-process.

After 41 or 64 hours, the fermentation broth must be extracted forrecovery safracin B or KCN treatment in the clarified broth for recoverysafracin B—cyano.

Example B

Obtention of safracin B cyano from the crude extract.

A clarification or filtration from the fermentation broth at pH 6removes the solids. The clarified broth was adjusted a pH 9.5 withdiluted sodium hydroxide and extracted twice with 2:1 (v/v) ethylacetate, methylene chloride or butyl acetate. The extraction was carriedout into an agitated-vessel during 20′, the temperature of the mixturewas maintained at 8 to 10° C. The two phases were separated by aliquid-liquid centrifuge. The organic phase was dried with sodiumsulphate anhydrous or frozen and then filtered for removing ice. Thisorganic phase (ethyl acetate layer) was evaporated until obtention of anoil-crude extract.

Example C

Obtention of safracin B cyano from the clarified broth.

A clarification or filtration from the fermentation broth at pH 6removes the solids. The clarified broth was adjusted at pH 3.9 withconcentrated acetic acid. 0.5 grams per litre of KCN are added to theclarified broth an incubated at 20° C. during 1 hour with agitation.Then, the temperature was decreased at 15° C. and the pH was adjusted at9.5 with diluted sodium hydroxide and extracted with 2:1.5 (v/v) ethylacetate. The extraction was carried out into an agitated-vessel during20 minutes, the temperature of the mixture was maintained at 8 to 10° C.The two phases were separated by a liquid-liquid centrifuge. The organicphase was dried with sodium sulphate anhydrous. This organic phase(ethyl acetate layer) was evaporated until obtention of an oil-crudeextract. This extract was purified by flash column chromatography (SiO₂,gradient 20:1 to 10: to 5:1 ethyl acetate:methanol) to affordquantitatively compound 2 as a light yellow solid.

Rf: 0.55 (ethyl acetate:methanol 5:1); .t_(R)=19.9 min [HPLC, Delta PackC4, 5 μm, 300 A, 150×3 mm, λ=215 nm, flow=0.7 ml/min, temp=50° C.,grad.: CH₃CN-aq. NaOAc (10 mM) 85%–70% (20′)];

¹H NMR (300 Mhz, CDCl₃): δ 6.54 (dd, J₁=4.4 Hz, J₂=8.4 Hz, 1H), 6.44 (s,1H), 4.12 (d, J=2.4 Hz, 1H), 4.04 (d, J=2.4 Hz, 1H), 4.00 (s, 3H), 3.87(bs, 1H), 3.65 (ddd, J₁=1.5 Hz, J₂=8.7 Hz, J₃=9.9 Hz, 1H), 3.35 (br. D,J=8.4 Hz, 1H), 3.15–2.96 (m, 4H), 2.92 (q, J=7.2 Hz, 1H), 2.47 (d,J=18.3 Hz, 1H), 2.29 (s, 3H), 2.18 (s, 3H) 1.83 (s, 3H), 1.64 (ddd,J₁=2.7 Hz, J₂=11.1 Hz, J₃=14.1 Hz, 1H), 0.79 (d, J=7.2 Hz, 3H);

¹³C NMR (75 Mhz, CDCl₃): δ 186.0 (q), 175.9 (q), 156.2 (q), 146.8 (q),142.8 (q), 140.7 (q), 136.6 (q), 130.5 (q), 128.8 (q), 127.0 (q), 120.5(s), 117.4 (q), 116.5 (q), 60.8 (t), 60.4 (s), 58.7 (t), 56.2 (s), 55.7(s), 54.8 (s), 54.8 (s), 54.4 (s), 50.0 (s), 41.6 (t), 39.8 (d), 25.2(d), 24.4 (d), 21.2 (t), 15.5 (t), 8.4 (t).

ESI-MS m/z: Calcd for C₂₉H₃₅N₅O₆: 549.6. Found (M+Na)⁺: 572.3.

Example D

A medium (50 l) composed of dextrose (2%), mannitol (4%), dry brewer'syeast (2%), ammonium sulphate (1%), potassium secondary phosphate(0.04%), potassium chloride (0.8%), iron (III) chloride 6-hydrate(0.001%), L-tyrosine (0.1%), calcium carbonate (0.8%), poly-(propyleneglycol) 2000 (0.05%) and antifoam ASSAF 1000 (0.2%) was poured into ajar-fermentor with 75 l total capacity and, after sterilisation,inoculated with seed culture (2%) of A2-2 strain (FERM BP-14) andaerated cultivation under agitation was carried out at 27° C. to 24° C.for 64 hours (aeration of 75 l per minute and agitation from 350 to 500rpm). The pH was controlled by automatic feeding of diluted sulphuricacid from 27 hours to final process. A 2% mannitol was added from 16hours to final process. The cultured medium (45 l) thus obtained was,after removal of cells by centrifigation, adjusted to pH 9.5 withdiluted sodium hydroxide, extracted with 25 litres of ethyl acetatetwice. The mixture was carried out into an agitated-vessel at 8° C. for20 minutes. The two phases were separated by a liquid-liquid centrifuge.The organic phases were frozen at −20° C. and filtered for removing iceand evaporated ice and evaporated until obtention of a 40 goil-dark-crude extract. After introduction of the cyanide group andpurification, 3.0 grams of safracin B cyano were obtained.

Example E

A medium (50 l) composed of dextrose (2%), mannitol (4%), dry brewer'syeast (2%), ammonium sulphate (1%), potassium secondary phosphate(0.02%, potassium chloride (0.2%), Iron (III) chloride 6-hydrate(0.001%, L-tyrosine (0.1%), calcium carbonate (0.8%, poly-(propyleneglycol) 2000 (0.05%) and antifoam ASSAF 1000 (0.2%) was poured into ajar-fermentor with 75 l total capacity and, after sterilisation,inoculated with seed culture (2%) of A2-2 strain (FERM BP-14) andaerated cultivation under agitation was carried out at 27° C. to 24° C.for 41 hours (aeration of 75 l per minute and agitation from 350 to 500rpm). The pH was controlled by automatic feeding of diluted sulphuricacid from 28 hours to final process. A 1% mannitol was added from 16hours to final process. The cultured medium (45 l) thus obtained was,after removal of cells by centrifugation, adjusted to pH 3.9 with 200 mlof conc. acetic acid. 25 grams of potassium cyanide 97% were added andafter 1 hour of agitation at 20° C., the pH was adjusted to 9.5 with1500 ml of a solution 10% sodium hydroxide. Then, extracted with 35litres of ethyl acetate. The mixture was carried out into an agitatedvessel at 8° C. for 20 minutes. The two phases were separated by aliquid-liquid centrifuge. The organic phase was dried by sodium sulphateanhydrous and evaporated until obtention of a 60 g oil-dark-crudeextract.

After chromatography, 4.9 grams of safracin B cyano were obtained.

REFERENCES

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1. A compound of the formula:

wherein: R′ is selected from the group consisting of —CH₂—N(R^(a))₂ and—CH₂—OR^(a), where R^(a) is selected from the group consisting of H;alkyl-CO—; haloalkyl-CO—; cycloalkylalkyl-CO—; haloalkyl-O—CO—;arylalkyl-CO—; arylalkenyl-CO—; heteroaryl-CO—; alkenyl-CO—; alkeny; andamino acid acyl; R⁵ is —OR″, where R″ is selected from the groupconsisting of H; alkyl-CO—; cycloalkyl—CO—; and haloalkyl-CO—; R¹⁸ is—OR, where R is selected from the group consisting of H, alkyl-CO—; andcycloalkylalkyl—CO—; and R²¹ is —OH; or a pharmaceutically acceptablesalt thereof.
 2. The compound of claim 1, which is of the formula:

wherein R¹, R⁵, R¹⁸, and R²¹ are as defined in claim
 1. 3. The compoundof claim 1, wherein R¹ is —CH₂—NHR^(a).
 4. The compound of claim 1,wherein R^(a) is -aa-R^(b) where aa is amino acid acyl and R^(b) is asdefined for R^(a).
 5. The compound of claim 4, wherein said amino acidacyl is further substituted with at least one R^(a) group.
 6. Thecompound of claim 4, wherein R¹ is —CH₂—NH-aa-R^(b) where aa is an aminoacid and R^(b) is selected from the group consisting of hydrogen;arylalkenyl-CO—; haloalkyl-CO—; alkyl-CO—; arylalkyl-CO—; and amino acidacyl.
 7. The compound of claim 6, wherein R¹ is —CH₂—NH-aa-R^(b) whereaa is alanine and R^(b) is selected from the group consisting ofhydrogen, CF₃CO—, trifluoromethylcinnamoyl, cinnamoyl, C₃F₇CO—, butyryl,3-chloroproprionoyl, hydrocinnamoyl, hexanoyl, phenylacetyl, and acetyl.8. The compound of claim 5, wherein R¹ is —CH₂—NR^(a)-aa-R^(b) where aais an amino acid, R^(a) is alkyl-CO— and R^(b) is haloalkyl-CO—.
 9. Thecompound of claim 8, wherein R¹ is —CH₂—NR^(a)-aa-R^(b) where aa isacetylalanine, R^(a) is selected from the group consisting of acetyl andbutyryl, and R^(b) is CF₃—CO—.
 10. The compound of claim 3, wherein R¹is —CH₂—NHR^(a) where R^(a) is selected from the group consisting ofhydrogen, alkyl-CO—; alkenyl-CO—; arylalkenyl-CO—; arylalkyl-CO—;heteroaryl-CO—; cycloalkylalkyl-CO—; and alkenyl.
 11. The compound ofclaim 10, wherein R¹ is —CH₂—NHR^(a) where R^(a) is selected from thegroup consisting of hydrogen, acetyl, isovaleroyl, decanoyl, cinnamoyl,hydrocinnamoyl, phenylacetyl, propionyl, myristoyl, stearoyl, hexanoyl,crotonyl, chloronicotinoyl, cyclohexylacetyl, cyclohexylpropionyl andallyl.
 12. The compound of claim 1, wherein R′ is —CH₂—OR^(a) whereR^(a) is selected from the group consisting of hydrogen; alkyl-CO—;arylalkyl-CO—; and arylalkenyl-CO—.
 13. The compound of claim 12,wherein R¹ is —CH₂—OR^(a) where R^(a) is selected from the groupconsisting of hydrogen; butyryl; trifluoromethylcinnamoyl; cinnamoyl.14. The compound of claim 5, wherein R⁵ is —OR″, where R″ is selectedfrom the group consisting of H; alkyl-CO where the alkyl has an oddnumber of carbon atoms, and ω-cyclohexylalkyl-CO.
 15. The compound ofclaim 14, wherein R⁵ is —OCOCH₃.
 16. The compound of claim 15, whereinR¹ is —CH₂—NHR^(a) where R^(a) is selected from the group consisting ofhydrogen, acetyl, isovaleroyl, decanoyl, cinnamoyl, hydrocinnamoyl,phenylacetyl, propionyl, myristoyl, stearoyl, hexanoyl, crotonyl,chloronicotinoyl, cyclohexylacetyl, cyclohexylpropionyl and allyl. 17.The compound of claim 15, wherein R¹ is —CH₂—OR^(a) where R^(a) isselected from the group consisting of hydrogen; butyryl;trifluoromethylcinnamoyl; and cinnamoyl.
 18. The compound of claim 1,wherein R¹⁸ is —OR, where R is selected from the group consisting of Hand alkyl-CO.
 19. The compound of claim 18, wherein R¹⁸ is —OH.
 20. Thecompound of claim 19, wherein R⁵ is —OCOCH₃.
 21. The compound of claim19, wherein R¹ is —CH₂—NHR^(a) where R^(a) is selected from the groupconsisting of hydrogen, acetyl, isovaleroyl, decanoyl, cinnamoyl,hydrocinnamoyl, phenylacetyl, propionyl, myristoyl, stearoyl, hexanoyl,crotonyl, chloronicotinoyl, cyclohexylacetyl, cyclohexylpropionyl andallyl.
 22. A compound of a formula selected from the following generalstructures I, II and III:

wherein R′, X₂, R₁ and R₆ are each independently selected from thegroups defined below: R′ X₂ R₁ R₆ H OH OH OH CH₂CH═CH₂ OAc OAc COCH₂CH₃OCH₂CH═CH₂ OMOM COCH₂CH₂CH₃ OCOOCH₂CH═CH₂ OCOCH₂C₆H₁₁ CO(CH₂)₄CH₃ OCOCF₃OCOCH₂CH₂C₆H₁₁ CO(CH₂)₁₂CH₃ OCOCH₂Cl OCOCH₂CH₂CH₃ CO(CH₂)₁₆CH₃OCOCH₂CH₂Cl OCO(CH₂)₄CH₃ COCH₂C₆H₁₁ OCOCF₂CF₂CF₃ OCO(CH₂)₈CH₃COCH₂CH₂C₆H₁₁ OCO(CH₂)₁₆CH₃ COOCH₂CCl₃ COCH₂Ph COCH₂CH₂Ph COCH═CHCH₃COCH═CHPh COCH═CHArCF₃ COCH(CH₃)NHCOCH₂CH₂Ph CO—(S)—CH(CH₃)NHCOCF₃CO—(R)—CH(CH₃)NHCOCF₃ CO—(S)—CH(NHCbz)CH(CH₃)₂ Boc CSNHPh

or a pharmaceutically acceptable salt thereof.
 23. A compound which isselected from the following formulae:

or a pharmaceutically acceptable salt thereof.
 24. A compound of theformula:

or a pharmaceutically acceptable salt thereof.
 25. A compound of theformula:

or a pharmaceutically acceptable salt thereof.
 26. A pharmaceuticalcomposition comprising a compound of the formula:

wherein: R¹ is selected from the group consisting of —CH₂—N(R^(a))₂ and—CH₂—OR^(a), where R^(a) is selected from the group consisting of H;alkyl-CO—; haloalkyl-CO—; cycloalkylalkyl-CO—; haloalkyl-O-CO—;arylalkyl-CO—; arylalkenyl-CO—; heteroaryl-CO—; alkenyl-CO—; alkenyl andamino acid acyl; R⁵ is —OR″, where R″ is selected from the groupconsisting of H; alkyl-CO—; cycloalkyl-CO—; and haloalkyl-CO—; R¹⁸ is—OR, where R is selected from the group consisting of H, alkyl-CO—; andcycloalkylalkyl-CO—; and R²¹ is —OH; or a pharmaceutically acceptablesalt thereof; together with a pharmaceutical acceptable carrier.
 27. Thecomposition of claim 26, wherein said compound is of the formula:

wherein R¹, R⁵, R¹⁸, and R²¹ are as defined as in claim 26; or apharmaceutically acceptable salt thereof.
 28. The composition of claim26, wherein R¹ is —CH₂—NHR^(a).
 29. The composition of claim 26, whereinR^(a) is -aa-R^(b) where aa is amino acid acyl and R^(b) is as definedfor R^(a).
 30. The composition of claim 29, wherein said amino acid acylis further substituted with at least one R^(a) group.
 31. Thecomposition of claim 29, wherein R¹ is —CH₂—NH-aa-R^(b) where aa is anamino acid and R^(b) is selected from the group consisting of hydrogen;arylalkenyl-CO—; haloalkyl-CO—; alkyl-CO—; arylalkyl-CO—; and amino acidacyl.
 32. The composition of claim 31, wherein R¹ is —CH₂—NH-aa-R^(b)where aa is alanine and R^(b) is selected from the group consisting ofhydrogen, CF₃CO—, trifluoromethylcinnamoyl, cinnamoyl, C₃F₇CO—, butyryl,3-chloroproprionoyl, hydrocinnamoyl, hexanoyl, phenylacetyl, and acetyl.33. The composition of claim 30, wherein R¹ is —CH₂—NR^(a)-aa-R^(b)where aa is an amino acid, R^(a) is alkyl-CO— and R^(b) ishaloalkyl-CO—.
 34. The composition of claim 33, wherein R¹ is—CH₂—NR^(a)-aa-R^(b) where aa is acetylalanine, R^(a) is selected fromthe group consisting of acetyl and butyryl and R^(b) is CF₃—CO—.
 35. Thecomposition of claim 28, wherein R¹ is —CH₂—NHR^(a) where R^(a) isselected from the group consisting of hydrogen, alkyl-CO—; alkenyl-CO—;arylalkenyl-CO—; arylalkyl-CO—; heteroaryl-CO—; cycloalkylalkyl-CO—; andalkenyl.
 36. The composition of claim 35, wherein R¹ is —CH₂—NHR^(a)where R^(a) is selected from the group consisting of hydrogen, acetyl,isovaleroyl, decanoyl, cinnamoyl, hydrocinnamoyl, phenylacetyl,propionyl, myristoyl, stearoyl, hexanoyl, crotonyl, chloronicotinoyl,cyclohexylacetyl, cyclohexyipropionyl and allyl.
 37. The composition ofclaim 26, wherein R′ is —CH₂—OR^(a) where R^(a) is selected from thegroup consisting of hydrogen; alkyl-CO—; arylalkyl-CO—; andarylalkenyl-CO—.
 38. The composition of claim 37, wherein R¹ is—CH₂—OR^(a) where R^(a) is selected from the group consisting ofhydrogen; butyryl; trifluoromethylcinnamoyl; and cinnamoyl.
 39. Thecomposition of claim 26, wherein R⁵ is —OR″, where R″ is selected fromthe group consisting of H; alkyl-CO where the alkyl has an odd number ofcarbon atoms, and ω-cyclohexylalkyl—CO—.
 40. The composition of claim39, wherein R⁵ is —OCOCH₃.
 41. The composition of claim 40, wherein R¹is —CH₂—NHR^(a) where R^(a) is selected from the group consisting ofhydrogen, acetyl, isovaleroyl, decanoyl, cinnamoyl, hydrocinnamoyl,phenylacetyl, propionyl, myristoyl, stearoyl, hexanoyl, crotonyl,chloronicotinoyl, cyclohexylacetyl, cyclohexylpropionyl and allyl. 42.The composition of claim 40, wherein R¹ is —CH₂—OR^(a) where R^(a) isselected from the group consisting of hydrogen; butyryl;trifluoromethylcinnamoyl; and cinnamoyl.
 43. The composition of claim26, wherein R¹⁸ is —OR, where R is selected from the group consisting ofH and alkyl-CO—.
 44. The composition of claim 43, wherein R¹⁸ is —OH.45. The composition of claim 44, wherein R⁵ is —OCOCH₃.
 46. Thecomposition of claim 44, wherein R¹ is —CH₂—NHR^(a) where R^(a) isselected from the group consisting of hydrogen, acetyl, isovaleroyl,decanoyl, cinnamoyl, hydrocinnamoyl, phenylacetyl, propionyl, myristoyl,stearoyl, hexanoyl, crotonyl, chloronicotinoyl, cyclohexylacetyl,cyclohexylpropionyl and allyl.
 47. The composition of claim 44, whereinR¹ is —CH₂—OR^(a) where R^(a) is selected from the group consisting ofhydrogen; butyryl; trifluoromethylcinnamoyl; and cinnamoyl.
 48. Acomposition comprising a compound of a formula selected from thefollowing general structures I, II and III:

wherein R′, X₂, R₁ and R₆ are each independently selected from thegroups defined below: R′ X₂ R₁ R₆ H OH OH OH CH₂CH═CH₂ OAc OAc COCH₂CH₃OCH₂CH═CH₂ OMOM COCH₂CH₂CH₃ OCOOCH₂CH═CH₂ OCOCH₂C₆H₁₁ CO(CH₂)₄CH₃ OCOCF₃OCOCH₂CH₂C₆H₁₁ CO(CH₂)₁₂CH₃ OCOCH₂Cl OCOCH₂CH₂CH₃ CO(CH₂)₁₆CH₃OCOCH₂CH₂Cl OCO(CH₂)₄CH₃ COCH₂C₆H₁₁ OCOCF₂CF₂CF₃ OCO(CH₂)₈CH₃COCH₂CH₂C₆H₁₁ OCO(CH₂)₁₆CH₃ COOCH₂CCl₃ COCH₂Ph COCH₂CH₂Ph COCH═CHCH₃COCH═CHPh COCH═CHArCF₃ COCH(CH₃)NHCOCH₂CH₂Ph CO—(S)—CH(CH₃)NHCOCF₃CO—(R)—CH(CH₃)NHCOCF₃ CO—(S)—CH(NHCbz)CH(CH₃)₂ Boc CSNHPh

or a pharmaceutically acceptable salt thereof; together with apharmaceutically acceptable carrier.
 49. A composition comprising acompound which is selected from the following formulae:

or a pharmaceutically acceptable salt thereof; together with apharmaceutically acceptable carrier.
 50. A composition comprising acompound of the formula

together with a pharmaceutically acceptable carrier; or apharmaceutically acceptable salt thereof.
 51. A composition comprising acompound of the formula:

together with a pharmaceutically acceptable carrier; or apharmaceutically acceptable salt thereof.
 52. A method of treating amammal affected by bladdar cancer, breast cancer, colon cancer, stomachcancer, liver cancer, lung cancer, ovarian cancer, pancreatic cancer,throat cancer, prostate cancer, kidney cancer, retinoblastoma, melanoma,fibrosarcoma, chondrosarcoma, osteosarcoma, leukemia, or lymphoma, whichcomprises administering to the affected mammal a therapeuticallyeffective amount of a compound of the formula:

wherein: R¹ selected from the group consisting of —CH₂—N(R^(a))₂ and—CH₂—OR^(a), where R^(a) is selected from the group consisting of H;alkyl-CO—; haloalkyl-CO—; cycloalkylalkyl-CO—; haloalkyl-O—CO—;arylalkyl-CO—; arylalkenyl-CO—; heteroaryl-CO—; alkenyl-CO—; alkeny; andamino acid acyl; R⁵ is —OR″, where R″ is selected from the groupconsisting of H; alkyl-CO—; cycloalkyl-CO—; and haloalkyl-CO—; R¹⁸ is—OR, where R is selected from the group consisting of H, alkyl-CO—; andcycloalkylalkyl-CO—; R²¹ is —OH; or a pharmaceutically acceptable saltthereof.
 53. The method of claim 52, wherein said mammal is a human. 54.The method of claim 53, wherein said compound is administered byintravenous infusion.
 55. The method of claim 52, wherein said compoundof formula:

is administered as part of a combination therapy.
 56. The method ofclaim 52, wherein said compound is of the formula:

wherein R¹, R⁵, R¹⁸, and R²¹ are as defined as in claim
 52. 57. Themethod of claim 52, wherein R¹ is —CH₂—NHR^(a).
 58. The method of claim52, wherein R^(a) is -aa-R^(b) where aa is amino acid acyl and R^(b) isas defined for R^(a).
 59. The method of claim 58, wherein said aminoacid acyl is further substituted with at least one R^(a) group.
 60. Themethod of claim 58, wherein R¹ is —CH₂—NH-aa-R^(b) where aa is an aminoacid and R^(b) is selected from the group consisting of hydrogen;arylalkenyl-CO—; haloalkyl-CO—; alkyl-CO—; arylalkyl-CO—; and amino acidacyl.
 61. The method of claim 60, wherein R¹ is —CH₂—NH-aa-R^(b) whereaa is alanine and R^(b) is selected from the group consisting ofhydrogen, CF₃CO—, trifluorocinnamoyl, cinnamoyl, C₃F₇CO—, butyryl,3-chloroproprionoyl, hydrocinnamoyl, hexanoyl, phenylacetyl, and acetyl.62. The method of claim 59, wherein R¹ is —CH₂—NR^(a)-aa-R^(b) where aais an amino acid, R^(a) is alkyl-CO— and R^(b) is haloalkyl-CO—.
 63. Themethod of claim 62, wherein R¹ is —CH₂—NR^(a)-aa-R^(b) where aa isacetylalanine, R^(a) is selected from the group consisting of acetyl andbutyryl and R^(b) is CF₃-CO—.
 64. The method of claim 57, wherein R¹ is—CH₂—NHR^(a) where R^(a) is selected from the group consisting ofhydrogen, alkyl-CO—; alkenyl-CO—; arylalkenyl-CO—; arylalkyl-CO—;heteroaryl-CO—; cycloalkylalkyl-CO—; and alkenyl.
 65. The method ofclaim 64, wherein R¹ is —CH₂—NHR^(a) where R^(a) is selected from thegroup consisting of hydrogen, acetyl, isovaleroyl, decanoyl, cinnamoyl,hydrocinnamoyl, phenylacetyl, propionyl, myristoyl, stearoyl, hexanoyl,crotonyl, chloronicotinoyl, cyclohexylacetyl, cyclohexyipropionyl andallyl.
 66. The method of claim 52, wherein R′ is —CH₂—OR^(a) where R^(a)is selected from the group consisting of hydrogen; alkyl-CO—;arylalkyl-CO—; and arylalkenyl—CO—.
 67. The method of claim 66, whereinR¹ is —CH₂—OR^(a) where R^(a) is selected from the group consisting ofhydrogen; butyryl; trifluoromethylcinnamoyl; and cinnamoyl.
 68. Themethod of claim 52, wherein R⁵ is —OR″, where R″ is selected from thegroup consisting of H; alkyl-CO where the alkyl has an odd number ofcarbon atoms, and ω-cyclohexylalkyl-CO—.
 69. The method of claim 68,wherein R⁵ is —OCOCH₃.
 70. The method of claim 69, wherein R¹ is—CH₂—NHR^(a) where R^(a) is selected from the group consisting ofhydrogen, acetyl, isovaleroyl, decanoyl, cinnamoyl, hydrocinnamoyl,phenylacetyl, propionyl, myristoyl, stearoyl, hexanoyl, crotonyl,chloronicotinoyl, cyclohexylacetyl, cyclohexylpropionyl and allyl. 71.The method of claim 69, wherein R¹ is —CH₂—OR^(a) where R^(a) isselected from the group consisting of hydrogen; butyryl;trifluoromethylcinnamoyl; and cinnamoyl.
 72. The method of claim 52,wherein R¹⁸ is —OR, where R is selected from the group consisting of H,and alkyl-CO—.
 73. The method of claim 72, wherein R¹⁸ is —OH.
 74. Themethod of claim 73, wherein R⁵ is —OCOCH₃.
 75. The method of claim 74,wherein R¹ is —CH₂—NHR^(a) where R^(a) is selected from the groupconsisting of hydrogen, acetyl, isovaleroyl, decanoyl, cinnamoyl,hydrocinnamoyl, phenylacetyl, propionyl, myristoyl, stearoyl, hexanoyl,crotonyl, chloronicotinoyl, cyclohexylacetyl, cyclohexyipropionyl andallyl.
 76. The method of claim 73, wherein R¹ is —CH₂—OR^(a) where R^(a)is selected from the group consisting of hydrogen; butyryl;trifluoromethylcinnamoyl; and cinnamoyl.
 77. A method of treating amammal affected by bladdar cancer, breast cancer, colon cancer, stomachcancer, liver cancer, lung cancer, ovarian cancer, pancreatic cancer,throat cancer, prostate cancer, kidney cancer, retinoblastoma, melanoma,fibrosarcoma, chondrosarcoma, osteosarcoma, leukemia, or lymphoma, whichcomprises administering to the affected mammal a therapeuticallyeffective amount of a compound selected from the following generalstructures I, II and III:

wherein R′, X₂, R₁ and R₆ are each independently selected from thegroups defined below: R′ X₂ R₁ R₆ H OH OH OH CH₂CH═CH₂ OAc OAc COCH₂CH₃OCH₂CH═CH₂ OMOM COCH₂CH₂CH₃ OCOOCH₂CH═CH₂ OCOCH₂C₆H₁₁ CO(CH₂)₄CH₃ OCOCF₃OCOCH₂CH₂C₆H₁₁ CO(CH₂)₁₂CH₃ OCOCH₂Cl OCOCH₂CH₂CH₃ CO(CH₂)₁₆CH₃OCOCH₂CH₂Cl OCO(CH₂)₄CH₃ COCH₂C₆H₁₁ OCOCF₂CF₂CF₃ OCO(CH₂)₈CH₃COCH₂CH₂C₆H₁₁ OCO(CH₂)₁₆CH₃ COOCH₂CCl₃ COCH₂Ph COCH₂CH₂Ph COCH═CHCH₃COCH═CHPh COCH═CHArCF₃ COCH(CH₃)NHCOCH₂CH₂Ph CO—(S)—CH(CH₃)NHCOCF₃CO—(R)—CH(CH₃)NHCOCF₃ CO—(S)—CH(NHCbz)CH(CH₃)₂ Boc CSNHPh

or a pharmaceutically acceptable salt thereof.
 78. A method of treatinga mammal affected by bladdar cancer, breast cancer, colon cancer,stomach cancer, liver cancer, lung cancer, ovarian cancer, pancreaticcancer, throat cancer, prostate cancer, kidney cancer, retinoblastoma,melanoma, fibrosarcoma, chondrosarcoma, osteosarcoma, leukemia, orlymphoma, which comprises administering to the affected mammal atherapeutically effective amount of a compound which is selected fromthe following formulae:

or a pharmaceutically acceptable salt thereof.
 79. A method of treatinga mammal affected by bladdar cancer, breast cancer, colon cancer,stomach cancer, liver cancer, lung cancer, ovarian cancer, pancreaticcancer, throat cancer, prostate cancer, kidney cancer, retinoblastoma,melanoma, fibrosarcoma, chondrosarcoma, osteosarcoma, leukemia, orlymphoma, which comprises administering to the affected mammal atherapeutically effective amount of a compound of the formula:

or a pharmaceutically acceptable salt thereof.
 80. A method of treatmenta mammal affected by bladdar cancer, breast cancer, colon cancer,stomach cancer, liver cancer, lung cancer, ovarian cancer, pancreaticcancer, throat cancer, prostate cancer, kidney cancer, retinoblastoma,melanoma, fibrosarcoma, chondrosarcoma, osteosarcoma, leukemia, orlymphoma, which comprises administering to the affected mammal atherapeutically effective amount of a compound of the formula:

or a pharmaceutically acceptable salt thereof.
 81. The compound of claim19, wherein R¹ is —CH₂—OR^(a) where R^(a) is selected from the groupconsisting of hydrogen; butyryl; trifluoromethylcinnamoyl; andcinnamoyl.
 82. The method of claim 52, wherein the cancer is leukemia,lung cancer, colon cancer, melanoma, kidney cancer, prostate cancer,lymphoma, breast cancer, or ovarian cancer.
 83. The method of claim 77,wherein the cancer is leukemia, lung cancer, colon cancer, melanoma,kidney cancer, prostate cancer, lymphoma, breast cancer, or ovariancancer.
 84. The method of claim 78, wherein the cancer is leukemia, lungcancer, colon cancer, melanoma, kidney cancer, prostate cancer,lymphoma, breast cancer, or ovarian cancer.
 85. The method of claim 79,wherein the cancer is leukemia, lung cancer, colon cancer, melanoma,kidney cancer, prostate cancer, lymphoma, breast cancer, or ovariancancer.
 86. The method of claim 80, wherein the cancer is leukemia, lungcancer, colon cancer, melanoma, kidney cancer, prostate cancer,lymphoma, breast cancer, or ovarian cancer.
 87. A compound of a formulaselected from the following general structures I, II and III:

wherein R′, X₂, R₁ and R₆ are each independently selected from thegroups defined below: R′ X₂ R₁ R₆ COCH₂CH₂CH₃ OH OH OH COCH═CHPh OAc OAcCOCH═CHArCF₃ OCH₂CH═CH₂ OMOM CO—(S)—CH(CH3)NHCOCF₃ OCOOCH₂CH═CH₂OCOCH₂C₆H₁₁ OCOCF₃ OCOCH₂CH₂C₆H₁₁ OCOCH₂Cl OCOCH₂CH₂CH₃ OCOCH₂CH₂ClOCO(CH₂)₄CH₃ OCOCF₂CF₂CF₃ OCO(CH₂)₈CH₃ OCO(CH₂)₁₆CH₃

or a pharmaceutically acceptable salt thereof.
 88. A compound which isselected from the following formulae:

or a pharmaceutically acceptable salt thereof.
 89. A compositioncomprising a compound of a formula selected from the following generalstructures I, II and III:

wherein R′, X₂, R₁ and R₆ are each independently selected from thegroups defined below: R′ X₂ R₁ R₆ COCH₂CH₂CH₃ OH OH OH COCH═CHPh OAc OAcCOCH═CHArCF₃ OCH₂CH═CH₂ OMOM CO—(S)—CH(CH3)NHCOCF₃ OCOOCH₂CH═CH₂OCOCH₂C₆H₁₁ OCOCF₃ OCOCH₂CH₂C₆H₁₁ OCOCH₂Cl OCOCH₂CH₂CH₃ OCOCH₂CH₂ClOCO(CH₂)₄CH₃ OCOCF₂CF₂CF₃ OCO(CH₂)₈CH₃ OCO(CH₂)₁₆CH₃

or a pharmaceutically acceptable salt thereof; together with apharmaceutically acceptable carrier.
 90. A composition comprising acompound which is selected from the following formulae:

or a pharmaceutically acceptable salt thereof; together with apharmaceutically acceptable carrier.
 91. A method of treating a mammalaffected by bladdar cancer, breast cancer, colon cancer, stomach cancer,liver cancer, lung cancer, ovarian cancer, pancreatic cancer, throatcancer, prostate cancer, kidney cancer, retinoblastoma, melanoma,fibrosarcoma, chondrosarcoma, osteosarcoma, leukemia, or lymphoma, whichcomprises administering to the affected mammal a therapeuticallyeffective amount of a compound selected from the following generalstructures I, II and III:

wherein R′, X₂, R₁ and R₆ are each independently selected from thegroups defined below; R′ X₂ R₁ R₆ COCH₂CH₂CH₃ OH OH OH COCH═CHPh OAc OAcCOCH═CHArCF₃ OCH₂CH═CH₂ OMOM CO—(S)—CH(CH3)NHCOCF₃ OCOOCH₂CH═CH₂OCOCH₂C₆H₁₁ OCOCF₃ OCOCH₂CH₂C₆H₁₁ OCOCH₂Cl OCOCH₂CH₂CH₃ OCOCH₂CH₂ClOCO(CH₂)₄CH₃ OCOCF₂CF₂CF₃ OCO(CH₂)₈CH₃ OCO(CH₂)₁₆CH₃

or a pharmaceutically acceptable salt thereof.
 92. A method of treatinga mammal affected by bladdar cancer, breast cancer, colon cancer,stomach cancer, liver cancer, lung cancer, ovarian cancer, pancreaticcancer, throat cancer, prostate cancer, kidney cancer, retinoblastoma,melanoma, fibrosarcoma, chondrosarcoma, osteosarcoma, leukemia, orlymphoma, which comprises administering to the affected mammal atherapeutically effective amount of a compound which is selected fromthe following formulae:

or a pharmaceutically acceptable salt thereof.